Contents / İçindekiler

Contents / İçindekiler
Erol Kodak, N.Münevver Pinar, Nezaket Adigüzel and Aydan Acar
POLLEN MORPHOLOGY OF SOME TAXA OF GENUS TANACETUM L.
(ASTERACEAE) IN TURKEY
TÜRKİYE TANACETUM L. CİNSİNİN BAZI TAKSONLARININ POLEN
MORFOLOJİSİ
2-10
Esra Demir
POLLEN ANALYSIS OF HONEY SAMPLES COLLECTED FROM KOMATİ
(ÇAMLIHEMŞİN) PLATEAU
KOMATİ (ÇAMLIHEMŞİN) YAYLASI BALLARINDA POLEN ANALİZİ
11-16
Fatih Dikmen and Ahmet Murat Aytekin
NOTES ON ROPHITES ALGIRUS PÉREZ, 1895 AND ROPHITES
QUINQUESPINOSUS SPINOLA, 1808 OF MEDITERRANEAN TURKEY
WITH AN UPDATED LIST OF SUBFAMILY ROPHITINAE
(HYMENOPTERA: HALICTIDAE) OF TURKEY
TÜRKİYE’DEKİ ROPHITINAE (HYMENOPTERA: HALICTIDAE)
ALTFAMILYASININ GÜNCEL LİSTESİ İLE TÜRKİYE’NİN AKDENIZ
HAVZASINDAKİ ROPHITES ALGIRUS PÉREZ, 1895 AND ROPHITES
QUINQUESPINOSUS SPINOLA, 1808 ÜZERINE NOTLAR
17-26
Ömür Gençay Çelemli, Kadriye Sorkun, Bekir Salih
CHEMICAL COMPOSITION OF PROPOLIS SAMPLES COLLECTED FROM
TEKIRDAG-TURKEY
TEKİRDAĞ-TÜRKİYE’DEN TOPLANAN PROPOLİS ÖRNEKLERİNİN
KİMYASAL İÇERİĞİ
28-32
MELLIFERA
12-24:2-10 (2012) HARUM
RESEARCH ARTICLE
2
POLLEN MORPHOLOGY OF SOME TAXA OF GENUS
TANACETUM L. (ASTERACEAE) IN TURKEY
TÜRKİYE TANACETUM L. CİNSİNİN BAZI TAKSONLARININ POLEN MORFOLOJİSİ
Erol Kodak*, N.Münevver Pinar*, Nezaket Adigüzel** and Aydan Acar*
Summary:Morphological features of pollen of 8 Turkish taxa of the complex genus Tanacetum L. were
examined using light (LM) and scanning electron (SEM) microscopy. The pollen is tricolporate, trisyncolporate or tricolpate. The shape is oblate- spheroidal. The exine is echinate. The ornamentations
between spines are granulate, reticulate and rugulate-granulate. The results indicate that the value of
pollen characters for taxonomic applications is limited for Tanacetum.
Keywords: Asteraceae, Tanacetum, Pollen, LM, SEM, systematics, melissopalynology.
Özet: Kompleks bir cins olan Tanacetum L. cinsinin Türkiye’deki 8 taksonun polenlerinin morfolojik
özellikleri ışık (LM) ve taramalı elektron (SEM) mikroskopları kullanılarak çalışılmıştır. Polenlerin
trikolporat, trisinkolporat ve trikolpat oldukları bulunmuştur. Polen şekli oblat-speroidal’dir. Ekzin
ekinat’dır. Spinler arasındaki ornamentasyonlar granulat, retikülat ve rugulat- granulat’tır. Sonuçlar,
taksonomik uygulamalar için polen karakterlerinin değerinin Tanacetum cinsi söz konusu olduğunda
sınırlı olduğunu işaret etmektedir.
Anahtar Kelimeler: Asteraceae, Tanacetum, Polen, LM, SEM, sistematik, melissopalinoloji.
* Ankara University Faculty of Science, Department of Biology, Tandoğan 06100 Ankara, TURKEY
**Gazi University Faculty of Science, Department of Biology, Ankara, TURKEY
Corresponding Author E-mail: [email protected]
3
Introduction
cetum is represented by 60 of which are 27 taxa (47%)
Bees require large amounts of pollen for their own endemic (Grierson 1975; Yildirimli 1989; Ekim et al.
reproduction. The general view of pollen as an easy- 2000).
to-use protein source for flower visitors. That is Apis
World
(Heywood
and
Humphries
1977; from
SorengIn and
Cope
1991).
In to
Turkey,
is
mellifera
L. feed on
pollen
and nectar collected
this study,
8 taxa
belonging
the genusTanacetum
Tanacetum
blooming flowers. Tanacetum L. is also visited by Apis were investigated; T. balsamita L. subsp. balsamita
represented
bypollen
60 ofdiets
which
are 27et taxa
(47%)
(Grierson
1975;
Yildirimli
1989;
mellifera for
(Christophe
al. 2008;
Hilty endemic
L., T. balsamita
L. subsp.
balsamitoides
(Schultz
Bip.)Ekim
Knowledge about the pollen morphology of ho- Grierson, T. argenteum (Lam.) Willd. subsp. flabelet al.2012).
2000).
ney plants is important in the identification of plant lifolium (Boiss. End Heldr.) Grierson, T. argenteum
species
which8contribute
toward composition
honey Tanacetum
(Lam.) Willd.were
subsp. investigated;
argenteum (L.) All.,
argente- L.
In this
study,
taxa belonging
to the ofgenus
T. T.balsamita
(Howes 1953; Sodré et al. 2001). Silici and Gökçeoglu um (Lam.) Willd. subsp. canum (C. Koch) Grierson, T.
(2007)
have presented
is an important
(Post) (Schultz
Grierson, T.Bip.)
haradjanii
(Rech. T.
subsp.
balsamita
L.,that
T.Tanacetum
balsamita
L. subsp. depauperatum
balsamitoides
Grierson,
bee plant in the Mediterranean region of Anatolia. T. Fil.) Grierson and T. tomentellum (Boiss.) Grierson. T.
argenteum
(Lam.)
flabellifolium
(Boiss. End
Grierson,
T. argenteum
vulgare L.
has beenWilld.
reportedsubsp.
as a minor
element in argenteum
subsp.Heldr.)
flabellifolium,
T. argenteum
subsp.
honey by Sorkun (2008). The genus Tanacetum is one argenteum, T. depauperatum, T. haradjanii are local
(Lam.)
Willd. subsp. argenteum (L.) All., T. argenteum (Lam.) Willd. subsp. canum (C.
of the more than 100 genera in the tribe Anthemideae endemic species. T. balsamita L. subsp. balsamita L.,
(Soreng
and Cope
which contains (Post)
about 10%
of T. balsamita
L. subsp.(Rech.
balsamitoides
(Schultz Bip.)
Koch)
Grierson,
T.1991)
depauperatum
Grierson,
T. haradjanii
Fil.) Grierson
and T.
the total genera and 15% of the species of Asteraceae Grierson, T. argenteum (Lam.) Willd. subsp. canum
(Heywood and
Humphries
1977). About
species (C.
Koch) Grierson
and T. tomentellum
(Boiss.) Gritomentellum
(Boiss.)
Grierson.
T. 150
argenteum
subsp.
flabellifolium,
T. argenteum
subsp.
of Tanacetum are spreaded around the world. They are erson are widely distributed, nonendemic taxa. The
argenteum,
T. depauperatum,
T. haradjanii
are distribution
local endemic
balsamita
found throughout
temperate, regions,
particularly in
map of species.
the taxa is T.
given
in Figure 1.L. subsp.
the northern hemisphere even up to Northern Europe,
balsamita
T. balsamita
subsp.
balsamitoides
(Schultz has
Bip.)
Grierson,
T. argenteum
Canada, L.,
Alaska,
and Northern L.
Russia
(Hultén
1950; Pollen morphology
provided
an approach
to the
Hultén 1968; Heywood 1976; Heywood and Humphri- systematic relationships among the genera of Astera(Lam.) Willd. subsp. canum (C. Koch) Grierson and T. tomentellum (Boiss.) Grierson are
es 1977), although the center of diversity and probably ceae (Wagenitz 1955; Stix 1960; Erdtman 1969; Pinar
alsodistributed,
the origin for Tanacetum
is South-West
Asiadistribution
and and Inceoglu
1996;
Adigüzel
Pinar
widely
nonendemic
taxa. The
map of
thePinar
taxaand
is given
in1998;
Figure
1.
the Caucasus in the Old World (Heywood and Hump- and Oybak Dönmez 2000; Punt and Hoen 2009). Thehries 1977; Soreng and Cope 1991). In Turkey, Tana- re are numerous publications on pollen morphology
Figure 1. Distribution of the eight investigated Tanacetum L. taxa in Turkey
Figure 1. Distribution of the eight investigated Tanacetum L. taxa in Turkey
Pollen morphology has provided an approach to the systematic relationships among the
MELLIFERA
of genus Tanacetum (İnceoğlu and Karamustafa 1977;
Ramos & Mederos 2008; Punt and Hoen 2009).
The aim of this study is to illustrate the range of variability in pollen characters of T. balsamita subsp. balsamita, T. balsamita subsp. balsamitoides, T. argenteum subsp. flabellifolium, T. argenteum subsp. argenteum, T. argenteum subsp. canum, T. depauperatum,
T. haradjanii, T. tomentellum found in Turkey in order
to establish their availability for future taxanomic and
melissopalynological works.
Material and Methods
The material was collected from wild populations.
The collectors and localities are provided in the “Specimens examined” for each taxon. The specimens are
deposited in GAZİ (Gazi University Herbarium), AEF
(Ankara University Farma Herbarium) and HUB (Hacettepe University Herbarium).
Pollen slides were prepared using by the technique
of Wodehouse (1935). LM studies were done with a
Leitz-Wetzlar microscope. Measurements are based
on at least 30 pollen grains for each taxon. For SEM
studies, pollen grains were coated with gold for four
minutes in a sputter-coater. Observations were made
with a Jeol 100 CXII electron microscope.
The pollen terminology follows Faegri-Iversen (1975)
and Punt et al. (1994). The Simpson and Roe graphical
test (Van der Pluym & Hideux 1977) was used for statistical calculations.
Specimens examined
The order of the species was adapted from Grierson
(1975). All the specimens are deposied in GAZİ, AEF
and HUB: T. balsamita L. subsp. balsamita L., Konya, Koyuncu AEF, Erzurum, Koyuncu AEF; T. balsamita L. subsp. balsamitoides (Schultz Bip.) Grierson, Hakkari Koyuncu AEF, Erzurum Koyuncu AEF,
Sivas Çelik AEF, Gümüşhane, Kars Koyuncu AEF;
T. argenteum (Lam.) Willd subsp. flabellifolium (Boiss. End Heldr.) Grierson, Konya Adigüzel GAZİ, T.
argenteum (Lam.) Willd. subsp. argenteum (L.) All
Kayseri Adigüzel GAZİ, Hakkari Koyuncu AEF; T.
argenteum (Lam.) Willd. subsp. canum (C. Koch) Grierson, Tunceli Adigüzel GAZİ, Içel Çelik AEF, Er-
4
zincan Soner AEF; T. depauperatum (Post) Grierson,
Hatay Adigüzel GAZİ; T. haradjanii (Rech.) Grierson
Adana Çelik AEF; T. tomentellum (Boiss.) Grierson
Bitlis, Peşmen HUB.
Results
Detailed pollen morphological features of the investigated taxa are summarized in Table 1 and Fig. 2 and
representative pollen grains are illustrated in Fig. 3-4.
Size, symmetry and shape
The pollen grains are radially symmetrical and isopolar. The shape is generally oblate-spheroidal (the
term according to Erdtman (1969) based on the P / E
ratio, Table 1, with polar axes ranging from 20.1 - 34
µm and equatorial axes from 25 to 34 µm, respectively. The largest pollen is found in T. balsamita subsp.
balsamita (chromosome number 4n = 36), while the
smallest pollen occurs in T. haradjanii. The outline
is circular or subcircular in equatorial view and generally triangular-convex and sometimes circular or
circular-convex in polar view. Amb intersemiangular.
(Table 1, Fig. 2-4).
Apertures
Pollen grains of Tanacetum are operculate and usually
tricolporate or rarely trisyncolporate and tricolpate.
Some species have shown heteromorphic characteristics. For example; 2% tricolpate and 98% tricolporate in T. balsamita subsp. balsamita and T. argenteum
subsp. argenteum and 2% trisyncolporate and 98%
tricolporate in T. balsamita subsp. balsamitoides. Colpus, short or long (11- 18.5 µm) and broad (5-9 µm),
and ora lalongated. The highest values were observed in T. balsamita subsp. balsamita and T. argenteum subsp. canum. Margins distinct, regular and ends
acute in both of them. Colpus membrane more or less
granulate. (Table 1 and Fig. 3,4).
Exine
The stratified exine has an overall thickness which
ranges from 3 to 5.5 µm in. Ectexine is thicker than
endexine without no costae and no cavea. Intratectal
columellae very distinct under spines, but indistinct
interspinal region. The spines are commonly conical
with a broadened base and a tappered apical portion.
The spine length varies between 2- 4.5 µm The width
5
of spines varies between 2- 4.5 µm. The base of the
spines in almost all species studied has 3 or 4 irregular seriate perforations with larger holes which are
often found distally. Also, large or small cavities are
present. Number of cavities are 15-40. Intine is thick
(0.75-1.5 µm) (Table 1 and Fig. 3,4).
The pollen wall is provided with spines and its either
granulate, reticulate and rugulate-granulate. In all of
the species, the tectum surrounding the spine base is
microperforate. Table 1 provides a summary of the
tectal morphology.
Discussion
The grains of taxa of Tanacetum can be ascribed to
the ‘’Anthemis type‘’ of Stix (1960), and Anthemis arvensis type of Punt and Hoen (2009). The results of
our investigation show that the pollen dimensions (E
and, P / E ratio), the thickness of the exine and intine,
the shape of the polar and equatorial views and the
aperture type of the Turkish taxa of Tanacetum were
comparatively homogenous (Table 1 and Figure 2).
Ramos and Mederos (2008) and Özmen et al. (2009)
confirmed that the pollen morphologies of taxa of Tanacetum are homogenous.
Most Tanacetum species are diploid, having 2n= 2x=
18 with the basic chromosome number x=9. The chromosome count (2n=36) of T. balsamita subsp. balsamita is also the report of the tetraploid level (Watanable 2011; Inceer and Hayırlıoğlu 2012). Chatuverdi
et al. (1990) and Brochman (1992) report that pollen
grain size strongly correlates with the level of polyploidy. In this study, the relationship between ploidy
level or chromosome number and pollen size at the taxon level in T. balsamita subsp. balsamita is demonstrated statistically; higher ploidy levels correspond to
an increase in pollen grain size (Table 1 and Fig. 2).
Mesfin et al. (1995) said that the ornamentations between spines are important characters for Asteraceae. Ornamentations between spines are granulate in
Tanacetum balsamita subsp. balsamita, T. balsamita
subsp. balsamitoides and T. haradjanii, or reticulate
in T. tomentellum, and rugulate-granulate in T. deparatum, T. argenteum subsp. argenteum, T. argenteum
subsp. flabellifolium and T. argenteum subsp. canum.
(Table 1 and Fig. 4).
The general aperture form is tricolporate, but T. balsamita subsp. balsamita (2% tricolpate, 98% tricolporate), T. argenteum subsp. argenteum (2% tricolpate,
98% tricolporate) and T. balsamita subsp. balsamitoides (2% trisyncolporate and 98% tricolporate) show
considerable aperture type variation (Table 1 and Fig.
3,4). Variations in pollen size and aperture type have
been attributed to heteromorphy in pollen grains by
Nair and Kaul (1965) and Inceoglu (1973).
The taxa can be identificated by the sculpturing types
in this study. But the other results of polar and equatorial axes, pollen shape, exine and intine thickness are
generally similar to those taxa of the species.
P/E
0,9
0,9
0,87
0,93
Pollen
Shape
oblatespheraidal
oblatespheraidal
oblatespheraidal
oblatespheraidal
oblatespheraidal
oblatespheraidal
oblatespheraidal
oblatespheraidal
Taxa
T. balsamita subsp.
balsamita
T. balsamita subsp.
balsamitoide
Grierson
T. argenteum subsp.
flabellifolium
T. argenteum subsp.
argenteum
T. argenteum subsp.
canum
T. depauperatum
T. haradjanii
T. tomentellum
0,89
0,82
0,95
0,97
24
21,5
23
24
22,8
20,1
23,1
24,1
Max
27
25
26
28
25
34
27
33
Min
25,25
23,4
24
26,2
23,9
27,05
25,05
28,45
Mean
Polar axes (P) µm
Plg: Length of porus, Plt: Latitute of porus
25
25
25
27
25
25
25
25
Max
29
29
30
31
29
34
28
32
Min
27,2
27
27
29,2
27
29,5
26,5
28,5
Mean
Equatorial axes (E) µm
4,75
3,7
3,75
4
4,5
4
3,75
4,25
Exine
1,25
1,07
1
1,25
1,25
1,12
0,87
1
Intine
tricolporate
tricolporate
tricolporate
tricolporate
2% tricolpate
tricolporate
98% tricolporate
98% tricolporate
98% tricolporate
Aperture type
2% trisyncolporate
2% tricolpate
15
14
14
17
14
14,25
11,9
16,75
Clg
8
6,5
6
8
7,5
6
6
8
Clt
Colpus µm
8
6,5
6
8
7,5
6
6
8
Plg
8
6,5
6
8
7,5
6
6
8
Plt
Pore (P)µm
3,25
3,25
2,25
2,25
3,75
2,25
3,25
2,87
Length of
spine µm
3,25
3,75
2
2,25
4,25
3,12
4,62
4,17
Base of
spine
µm
Spine µm
Reticulate
Granulate
Rugulate Granulate
Rugulate Granulate
Rugulate Granulate
Rugulate Granulate
Granulate
Granulate
Ornamentation of
between spines
Table 1. The palynological mesurements and observations of the eight inventigated Tanacetum L. taxa. Clg: Length of colpus, Clt: Latitude of colpus,
MELLIFERA
6
sa
m
ba
l
sa
m
it a
T.
it o
id
es
f la
be
l li f
ol
iu
T.
m
ar
ge
nt
eu
m
T.
T.
ca
nu
de
m
pa
up
er
at
um
T.
ha
ra
dj
T.
an
to
ii
m
en
te
ll u
m
T.
ba
l
T.
Equatorial axes (E) µm
sa
m
ba
l
sa
m
it a
it o
id
es
f la
be
l li f
ol
iu
T.
m
ar
ge
nt
eu
m
T.
T.
ca
nu
de
m
pa
up
er
at
um
T.
ha
ra
dj
T.
an
to
ii
m
en
te
ll u
m
T.
T.
ba
l
T.
Polar axes (P) µm
7
40
35
30
25
Max
20
Min
15
Mean
10
5
0
Taxa
A
40
35
30
25
Max
20
Min
15
Mean
10
5
0
Taxa
B
Figure 2. A Polar axes (P), B Equatorial axes (E)
Figure 2. A Polar axes (P), B Equatorial axes (E)
MELLIFERA
8
A
B
C
D
E
F
G
H
I
K
L
M
N
O
P
S
T
U
W
X
Y
R
V
Z
Figure 3. LM photos of Tanacetum species. A-C: T. balsamita L. subsp. balsamita L. A. Polar view B. Polar view and
apertures C. Equatorial view. D-F: T. balsamita L. subsp. balsamitoides (Schultz Bip.) Grierson. D. Polar view. E. Polar
view and apertures. F. Equatorial view. G-I: T. argenteum (Lam.) Willd. subsp. flabellifolium (Boiss. End Heldr.) Grierson
G. Polar view. H. Polar view ornamentation. I. Equatorial view. K-M: T. argenteum (Lam.) Willd. subsp. argenteum (L.)
All.. K. Polar view. L. Polar view of ornamentation. M. Equatorial view. N-P: T. argenteum (Lam.) Willd. subsp. canum
(C. Koch) Grierson. N. Polar view O. Polar view and apertures P. Equatorial view. R-T: T. depauperatum (Post) Grierson.
R. Polar view. S. Polar view and apertures. T. Equatorial view. U-W: T. haradjanii (Rech. Fil.) Grierson U. Polar view.
V. Polar view ornamentation. W. Equatorial view. X-Z: T. tomentellum (Boiss.) Grierson. X Polar view. Y. Polar view and
apertures. Z. Equatorial view
9
A
B
C
D
E
F
G
H
I
K
L
M
N
O
P
R
S
T
U
V
W
X
Y
Z
Figure 4. SEM photos of Tanacetum species. A-C: T. balsamita L. subsp. balsamita L. A. Equatorial view B. Polar view
and apertures C. Ornamentation. D-F: T. balsamita L. subsp. balsamitoides (Schultz Bip.) Grierson. D. Equatorial view
E. Polar view and apertures F. Ornamentation. G-I: T. argenteum (Lam.) Willd. subsp. flabellifolium (Boiss. End Heldr.)
Grierson G. Equatorial view H. Polar view and apertures I. Ornamentation. K-M: T. argenteum (Lam.) Willd. subsp. argenteum (L.) All.. K. Equatorial view L. Polar view and apertures M. Ornamentation.. N-P: T. argenteum (Lam.) Willd.
subsp. canum (C. Koch) Grierson. N. Equatorial view O. Polar view and apertures P. Ornamentation.. R-T: T. depauperatum (Post) Grierson. R. Equatorial view S. Polar view and apertures T. Ornamentation.. U-W: T. haradjanii (Rech. Fil.)
Grierson U. Equatorial view V. Polar view and apertures W. Ornamentation.. X-Z: T. tomentellum (Boiss.) Grierson. X.
Equatorial view Y. Polar view and apertures Z. Ornamentation.
MELLIFERA
10
References
Özmen E., Kızılpınar İ., Özüdoğru B., Doğan C. and Erik S.
2009. Pollen morphology of some taxa of aromatic
genus Tanacetum L. (Asteraceae). Fabad J. Pharm.
Sci. 34, 1-11.
Chatuverdi M, Yunus D & Nair PK. 1990. Cytopalynological
studies of Arachis L. (Leguminosae). Cultivated and
wild species and their hybrids. Grana 29, 109-117.
Pınar N.M. and İnceoğlu Ö. 1996. A Comparative study on the
pollen morphology of Centaurea triumfettii All. groups A, B and C with light and electron microscopy.
Turkish Journal of Botany, 20, 395-398
Brochman C. 1992. Pollen and seed morphology of Nordic Draba (Brassicacea): phylogenetic and ecological implications. Nord J Bot, 12, 657-673.
Christophe J., Andreas M.,and Silvia D. 2008. Specıalızed bees
faıl to develop on non-host pollen: Do plants chemıcally protect theır pollen Ecology, 795–804
Ekim T, Koyuncu M, Vural M, Duman H, Aytac Z, Adıguzel
N. 2000. Türkiye Bitkileri Kırmızı Kitabı, Ankara:
Türkiye Tabiatını Koruma Derneği.
Erdtman G. 1969. Handbook of Palynology, Morphology, Taxonomy and Ecology. Munksgaard, Copenhagen.
Faegri K & Iverson J. 1992. Textbook of Pollen Analysis. 4th
Edition. New York: Wiley.
Grierson AJC. 1975. Tanacetum L. (emend. Briq.) in P.H. Davis
(Ed.) “Flora of Turkey and the East Aegean Islands”,
Edinburgh: Edinburgh University Press, 5, 256–292,
Heywood V.H. 1976. Tanacetum. In: Tutin, T.G., Heywood,
V.H., Burges, N.A., Moore, D.M.
Heywood V.H. and Humphries C.J. 1977. Anthemideae-systematic review. In: Heywood.
Hilty J. 2012. Insect Visitors of Illinois Wildflowers. World
Wide Web electronic publication. illinoiswildflowers. info, version.
Pınar N.M. and Adıgüzel N. 1998. Pollen morphology of some
Turkish Artemisia L. (Compositae) species. Ot Sistematik Botanik Dergisi, 5(2), 87-92.
Pınar N.M. and Oybak Dönmez E. 2000. Pollen morphology of
some Turkish endemic Helichrysum Gaertner species (Compositae). Pakistan Journal of Botany 32 (2),
295-301.
Punt W., Blackmore S., Nilson S. And Le Thomas A. 1994.
Glossary of polen an spore terminology LPP foundation, Utrecht.
Punt W & Hoen PP 2009. The Northwest European Pollen Flora,70: Asteraceae-Asteroideae. Rev Palaeobot and
Palyno 157, 22-183.
Ramos I.E. and Mederos M.A.P. 2008. Pollen morphology of
endemic species of the Gonospermum Less., Lugoa
DC. and Tanacetum L. complex
S. Silici & M. Keceoglu 2007. Pollen analysis of honeys from
Mediterranean region of Anatolia Grana; 46, 57–64
Howes F.N. 1953. Plantas melíferas. Barcelona, Espanha,
Reverté, 35p.
Sodré G.D., Marchini L.C., Moreti A.C. and Carvalho C. 2001.
Análises polínicas de méis de Apis mellifera L., 1758
(Hymenoptera: Apidae) do litoral norte do Estado da
Bahia. Rev Agric, 76, 215-225.
Hultén E. 1950. Atlas of the distribution of vascular plants in
NW Europe. Generalstabens Litografiska Anstalts
Förlag, Stockholm, 512 p.
Soreng R.J. and Cope E.A. 1991. On the taxonomy of cultivated species of the Chrysanthemum Genus-Complex
(Anthemideae; Compositae). Baileya, 23, 145-165.
Hultén E. 1968. Flora of Alaska and Neighboring territories.
A manual of the vascular plants.Stanford University
Press, Stanford, California. 1008 p.
Sorkun K. 2008. Türkiye’nin Nektarlı Bitkileri, Polenleri ve
Balları. Palme Yayıncılık, Ankara.
İnceoğlu Ö. and Karamustafa F. 1977. The pollen morphology
of plant in Ankara region I. Compositae. Communications 21, 77-100.
İnceer H. Hayırlıoglu Ayaz S. 2012. Karyological studies of
some representatives of Tanacetum L. (Anthemideae-Asteraceae) from North-east Anatolia. Plant.Syst.
Evol 298, 827-834.
İnceoğlu O. 1973. Asyneuma canescens (W. K.) Griseb. &
Schenk’in polen morfolojisi ve heteremof polenler.
Türk Biyoloji Derg 23, 89-94.
İrena E. La Serna Ramos and Miguel A.PadronMederos 2008.
Polen morphology of endemic species of the Gonospermum Less., Lugoa DC.
And Tanacetum L.complex (Asteraceae: Anthemideaae) in the
Canary Islands (Spain), and its taxonomical implications. Grana 47, 247-261.
Mesfin T., Crawford d.j. and Smith E.B.1995. Pollen morphology
of North Coreopsis (Compositae). Grana 34, 21-27
Nair PK & Kaul KN. 1965. Pollen grain in a gigantic of Rauwolfia serpentine. Current Sci 34, 256-257.
Stix E. 1960. Pollenmorphologische Untersuchungen an Compositen. Grana Palynol, 2(2), 41-114.
Watanable W. 2011. Index to chromosome numbers in Asteraceae. http://www.lib.kobe-u.ac.jp/infolib/meta_pub/
G0000003asteraceae_e, updated.
Wagenitz G. 1955. Pollenmorphologie und Systematik in der
Gattung Centaurea L. s.l. Flora, 142, 213-277.
Wodehouse PP 1935. Pollen Grains. New York: McGraw-Hill.
Valentine D.H., Walters S.M. and Webb D.A. (eds). Flora Europaea, Plantaginaceae to Compositae (and Rubiaceae).
Cambridge University Press, Cambridge, 4,169-171.
Van der Pluym A & Hideux M 1997. Applications d’une methodologie quantitative a la palynologie d’Eryngium
maritimum (Umbelliferae), Plant Syst Evol, 127, 5585.
Yıldırımlı Ş. 1989. Munzur dağlarının yeni, ilginc ve tukenen
bitki turleri. Hacettepe Fen ve MühendislikBilimleri
Dergisi, 10, 39-47,
Aytuğ B.1971. İstanbul Çevresi Bitkilerinin Polen Atlası. İstanbul Üniversitesi Orman Fakültesi
MELLIFERA
12-24:11-16 (2012) HARUM
RESEARCH ARTICLE
POLLEN ANALYSIS OF HONEY SAMPLES
COLLECTED FROM KOMATİ (ÇAMLIHEMŞİN)
PLATEAU
KOMATİ (ÇAMLIHEMŞİN) YAYLASI BALLARINDA POLEN ANALİZİ
Esra Demir*
Summary: This study presents the pollen analyses of 10 floral honeys from Komati (Çamlıhemşin) Plateau in Rize. Samples were obtained from each beehive selected randomly from 10 different apiaries in the
year 2011. All the honey samples were identificated under the light microscope. These samples examined
Total Pollen Number in 10 g honey (TPN-10g). The pollen analyses revealed 1 unifloral honey and 9 multifloral honeys. The dominant group of pollen grains consisted of : Castanea sativa MILLER., Ericaceae,
Fabaceae and Rosaceae. The amount of moisture in the samples of honey was identified between 15%
and 19%.
Keywords: Honey, Pollen Analysis, TPN-10g, Komati Plateau
Özet: Bu çalışma Komati Yaylası’ndan 10 bal örneğinin polen analizlerini içermektedir. Örnekler, 2011
yılında, rastgele belirlenmiş 10 farklı arılıktan seçilen birer kovandan toplanmıştır. Toplanan bütün bal
örnekleri ışık mikroskobu altında teşhis edilmişlerdir. Bu örneklerde Toplam Polen Sayısı (TPS-10) incelenmiştir. Polen analizleri sonucunda 1 bal unifloral ve 9 bal multifloral olarak saptanmıştır. Dominant
polen gruplarını Castanea sativa MILLER., Ericaceae, Fabaceae ve Rosaceae taksonları oluşturmaktadır. Bal örneklerindeki nem miktarı %15 ile %19 arasında tespit edilmiştir.
Anahtar Kelimeler: Bal, Polen Analizi, TPS-10g, Komati Yaylası
* Recep Tayyip Erdoğan University Faculty of Science, Department of Biology, 53100 Rize, TURKEY
Corresponding Author E-mail: [email protected]
11
MELLIFERA
Introduction
Honey is great importance for commercial and importance source of nutriment for people. The taste, smell
and color of honey is to change according to the nectar
of the flowers (Kaya et al., 2005).
Honey is the natural sweet substance produced by
honeybees from the nectar of blossoms or from secretions of living parts of plants or excretions which honeybee (Apis mellifera L.) collect, transform and combine with specific substances of their own, store and
leave in the honey comb to ripen and mature (Ünal
and Küplülü, 2006).
Beekeeping activity can be pursued only in such regions where bee flora is available. Therefore, identification of bee forage plants and their propagation help
in improving the bee forage wealth and the concomitant efficacy of beekeeping industry and commercial
honey production.
The melissopalynological studies have significant
application in the establishment of apiary industries.
Analyses of pollen from honey and pollen loads provide relevant information for the pollen and nectar
sources of an area. This knowledge is helpful for developing an apiary industry and commercial honey
production (Bhusari et al., 2005).
12
tained from Chubut (Forcone 2008), 13 samples obtained from Arıt (Mısır 2011), 20 samples obtained
from Burdur (Taşkın 2009).
Honey is hygroscopic; that is, it has excellent water absorbing properties. Thus honey will change in
moisture content according to the surrounding atmosphere. This characteristic is important in storing
honey because it will absorb water when exposed to
high relative humidity (RH) and will give off water
when exposed to low RH, until an equilibrium point is
reached. High moisture honey will ferment (Sanford
1994).
The amount of moisture that occurred in honey is a
very important factor and the criteria that determine
the quality of honey. The amount of honey is affected
by plant source, temperature, rains, condition of bee
glaze, the work during the marketing and degree of
maturation of honey. Keeping the honey not soured
is related to the amount of water in the structure of
honey. According to Turkish food Codex, Notification
of Honey, the amount of moisture of honey can not be
more than the value 20% (MEB, 2012).
The current study aimed at identifying nectar containing flower plants, which contribute to the formation of
honey from Komati Plateau of Çamlıhemşin in Rize.
The first pollen analyses of honey were studied by
Pfister (Kaya et al., 2005). In Turkey pollen analysis
of honey was the first made by Qustiani (Stawiartz
and Wreblewska, 2010). The first pollen analysis were
carried out by Sorkun and İnceoğlu between 1979 and
1981 (Çam et al., 2010).
Material and Method
In Turkey Başoğlu et al. (1996) were the first to determine honey quality by using the TPN-10 method.
From 1996 and on, quality assessment trough the
TPN-10 g as a melissopalynologic analysis has continued (Bölükbaşı, 2009).
The preparation of the honey samples were done using
the method defined by the International Bee Research
Association (Von Der Ohe et al., 2004). Preparations
were made from each honey samples for identification of pollen. After identification, 200 pollen samples were counted in each preparation. Source books
(Aytuğ, 1971) were used during the pollen analyses.
Olympus CX21 microscope was used for the analyses.
Pollen analysis were performed in 73 honey samples
obtained from Sandomierska (Stawiardz and Wreblewska, 2010), 78 samples obtained from Muğla (Özkök, 2009), 25 honey samples obtained from Canary
Islands (La-Serna Ramos et al. 1999), 14 samples ob-
The honey samples were collected from different beehive the month of July- August in 2011. During the
field studies, herbarium materials were collected.
Reference pollen preparations was made from the herbarium materials.
Pollen types were classified into four categories: between 1% and 5% was considered as the rare group,
13
between 6% and 20% was considered as the minor
group, between 21% and 50% was considered secondary group and pollen exceeding 50% was called the
dominant group (Doğan and Sorkun 2001). Based on
the total number of plant elements, honeys are placed
into one of the following classes (Sorkun 2008): Class
I with less than 2000 pollen grains per gram of honey
(includes unifloral honeys with underrepresented pollen); class II with 2000–10 000 pollen grains including most of multifloral and honeydew honeys and mixtures of flower and honeydew honeys; class III with 10
000–50 000 pollen grains includes unifloral honeys
with overrepresented pollen and honeydew honeys;
class IV with between 50 000–100 000 including
unifloral honey with strongly overrepresented pollen
and some pressed honeys; and class V with more than
100 000 pollen grains (Dobre et al. 2012).
The amount of moisture in honey samples which was
stored at 20°C was measured with Portable Refractometer.
Results
The results of melissopalynologic analyses, 9 samples
were identified multifloral honey, because they included pollen grains of multiple taxa. A sample which is
unifloral, referred as chesnut honey. In the honey samples 1,3,4,5,7 and 10 no pollen was found in dominant
amount. The sample number 6 Rosaceae pollen grains
were found in dominant amount, whereas Ericaceae
and Onobrychis pollen grains were found secondary
amount. In the honey sample 8 Ericaceae pollen grains
were found in dominant amount. In the honey sample
9 Castanea sativa MILLER. pollen grains were found
in dominant amount while no pollen grains are found
secondary and minor amount. Lauraceae and Poacaea
pollen grains were found rare amount in the sample
number 9 (Table I).
In this study ,TPN-10 g values ranged from 4 927 to 31
215. The sample number 9 contained the least number
of TPN-10 g and sample number 1 contained the most
number of TPN 10 (Table I).
Moisture content of the honey samples were up to
standart of honey notification of Turkish Food Codex
2012.
Discussion
The results of microscopic analyses revealed that taxa
variability is greatest in the rare group, followed by
minor, secondary and dominant groups (Table I). This
seems to confirm the view that variability is always
little amoung pollen taxa in dominant groups, while
greater among rare, minor and secondary groups
(Doğan and Sorkun 2001).
It is determined that nectar contributing to formation
of honey is obtained from plants with pollen grains
included in dominant and secondary groups (Erdoğan
et al. 2009).
In the study that made on the honey samples from
Komati Plateau pollen grains of Castanea sativa
MILLER. Ericaceae and Rosaceae were determined
in dominant group. In seven samples no pollen grains
were found in dominant group.
Castanea sativa MILLER. pollen grains were found to
be dominant due to the prevalence of chestnust trees.
In the multifloral honey samples were obtained pollen
grains in seconder and minor group while in the unifloral honey sample no pollen grains were obtained in
seconder and minor groups.
Acknowledgements
Techniqual support of this work by Kadriye Sorkun,
Ömür Gençay Çelemli and Şeyda Turan is gratefully
acknowledged.
MELLIFERA
14
Table I. Pollen Spectra, TPN-10 g Values (*Dominant pollen,**Secondary Pollen,***Minor Pollen,****Rare
Pollen) and the Amount of Moisture
H. S.
Number
1
2
3
4
5
6
7
8
9
10
Pollen spectrum
*
** Castanea sativa. MILLER., Ericaceae, Rosaceae
*** **** Brassicaceae, Fabaceae, Pinaceae, Rumex
*
** Ericaceae, Rosaceae
*** Castanea sativa MILLER., Fabaceae
**** Dipsacaceae, Lamiaceae, Rumex sp., Salix sp. , Tilia sp.
*
** Castanea sativa MILLER., Ericaceae, Rosaceae
*** Fabaceae
**** Asteraceae, Boraginaceae, Brassicaceae, Poaceae
*
** Castanea sativa MILLER., Ericaceae
*** Fabaceae, Rosaceae
**** Brassicaceae, Lamiaceae, Poaceae, Ranunculus sp.
*
** Castanea sativa MILLER.
*** Rosaceae
**** Ericaceae, Fabaceae, Salix sp.
*
Rosaceae
** Ericaceea, Onobrichis sp.
*** Castanea sativa MILLER.
**** Salix sp.
*
** Ericaceae, Rosaceae
*** Fabaceae, Rumex sp.
**** Brassicaceae, Lamiaceae, Pinaceae, Salix sp.
*
Ericaceae
** Rosaceae
*** Salix sp.
**** Cistus sp., Lamiaceae, Onobrychis sp., Rumex sp.
*
Castanea sativa MILLER.
** *** **** Lauraceae, Poaceae
*
** Castanea sativa MILLER., Ericaceae, Rosaceae
*** Fabaceae
**** Boraginaceae, Brassicaceae
TPN-10 g
31 215
18 898
27 123
26 272
9 585
9 531
26 728
10 500
4 927
12 542
The Amount
of Moisture
%
17.5
15
15.5
18.5
19
17
18
18.5
18.5
18
15
Table 2. Number of Pollen in Honey Samples
Sample No
Taxa
Number of
Pollen
Ericaceae
69
Castanea sativa MILLER.
Rosaceae
Fabaceae
1
2
3
4
5
Taxa
Number of
Pollen
Rosaceae
103
65
Ericaceae
47
58
Onobrychis sp.
28
Castanea sativa
MILLER.
19
6
Sample No
6
Brassicaceae
1
Salix sp.
3
Pinaceae
1
Ericaceae
85
Rumex sp.
1
Rosaceae
75
Ericaceae
95
Fabaceae
22
Rosaceae
57
Brassicaceae
9
7
Fabaceae
32
Salix sp.
5
Castanea sativa MILLER.
10
Lamiacaea
2
Lamiacaea
2
Pinaceae
1
Rumex sp.
2
Poaceaea
1
Tilia sp.
1
Ericaceae
106
Dipsacaceae
1
Rosaceae
66
Castanea sativa MILLER.
73
Salix sp.
22
Cistus sp.
2
Rumex sp.
2
15
Onobrychis sp.
1
Brassicaceae
3
Lamiaceae
1
Asteraceae
1
Castanea sativa
MILLER.
190
Poaceae
1
Poaceae
5
Boraginaceae
1
Lauraceaea
5
Castanea sativa MILLER.
90
Castanea sativa
MILLER.
80
Ericaceae
50
Rosaceae
60
Rosaceae
34
Ericaceae
41
Fabaceaea
12
Fabaceae
13
Brassicaceae
11
Boraginaceae
4
Lamiaceae
1
Ranunculus
1
Brassicaceae
1
Poaceae
1
Castanea sativa MILLER.
149
Rosaceae
40
Fabaceae
6
Ericaceae
3
Salix sp.
2
Rosaceae
64
Ericaceae
42
Fabaceae
8
9
10
MELLIFERA
16
Figure I. Microphotographs of Pollen Grains
a. Castanea sativa
MILLER.
b. Ericaceae
e. Rumex sp.
f.
1µ : x100
Lamiaceae
c. Cistaceae
d. Asteraceae
g. Pinaceae
h. Rosaceae
2µ : x40
Figure I. Microphotographs of Pollen Grains
References
Aytuğ
B. 1971. İstanbul Çevresi Bitkilerinin Polen
Atlası. İstanbul Üniversitesi Orman Fakültesi
Başoğlu F. N., Sorkun K., Löker M., Doğan C. and Wetherilt H.
1996. Saf ve Sahte Balların Ayırt Edilmesinde Fiziksel, Kimyasal ve Palinolojik Kriterlerin Saptanması.
Gıda. 21(2), 67-73.
Bhusari, N. V., Mate, D. M., Makde, K. H. 2005. Pollen of Apis
honey from Maharashtra. Grana. 44: 216–224. ISSN
0017-3134.
Bölükbası N. D., 2009. Melissopalynologıc Analysis of Pocked
Honey.Acknowledgements
Mellifera. 9-18:2-8.
Çam B., Pehlivan S., Uraz G., Doğan C.2010. Pollen Analyses of
Honey Collected from Various Regions of Ankara (Turkey) and Antibacterial Activity of These Honey Samples Against Some Bacteria. Mellifera. 10-19:2-16.
Dobre I., Alexe P., Escuredo O., Seıjo M. C. 2012. Palynological
evaluation of selected honeys from Romania. Grana.
iFirst: 1–9.
Doğan C., Sorkun K. 2001. Pollen Analysıs of Honeys From
Aegean, Marmara, Mediterranean and Black Sea
Regions İn Turkey. Mellifera.1-1: 33-34
Erdoğan N., Pehlivan S., Doğan C. 2009. Pollen Analysis
of Honeys from Sapanca-Karapürçek-Geyve and
Taraklı Districts of Adapazarı Province (Turkey).
Mellifrea. 9-17:9-18.
Forcone A. 2008. Pollen Analysis of Honey from Chubut (Argentinean Patagonia) Grana. 47: 147–158
Kaya Z., Binzet R., Orcan N. 2005. Pollen Analyses of Honeys
From Some Regions in Turkey Apiacata. 40:10-15
La-Serna Ramos, I. E., MeÂndez PeÂrez, B. & Gomez Ferreras, C. 1999. Pollen Characterization of Multiforal
Honeys from La Palma (Canary Islands). Grana 38:
356±363. ISSN 0017-3134.
Mısır M., 2011. Arıt Bölgesi (Bartın) Ballarında Polen Analizi,
Yüksek Lisans Tezi, Bartın.
Özkök A. 2009. The Microscopic, Organoleptic And Chemical
Analysis Of Pine Honey And Propolis, Which Are
Produced In Muğla Region. Doktora Tezi, Ankara.
Sanford T. M. 1994. Moisture in Honey.University of Florida
IFAS Extension.
Sorkun K. 2008. Türkiye`nin Nektarlı Bitkileri, Polenleri ve
Balları. Palme Yayıncılık.
Stawiardz E., Wroblewska A. 2010. Melissopalynolgıc Analysis
Of Multifloral Honeys From The Sandomierska Upland Area Of Poland. Journal of Apicultural Science.
65-75:54- 1
Taşkın, D., 2009. Burdur Yöresi Ballarının Polen Analizi, Yüksek Lisans Tezi, Isparta
Ünal C., Küplülü Ö.2006. Chemical Quality of Strained Honey
Consumed in Ankara. Ankara Üniv Vet Fak Derg,
53, 1-4.
Von Der Ohe W., Persano Oddo L., Piana M.L., Morlot M.,
Martin P.2004. Harmonized Methods of Melissopalynology. Apidologie. 35 S18–S25.
Laboratuvar Hizmetleri Bal Analizleri-1 524LT0028. 2012.
MEB, Ankara.
MELLIFERA
12-24:17-26 (2012) HARUM
RESEARCH ARTICLE
NOTES ON ROPHITES ALGIRUS PÉREZ, 1895 AND
ROPHITES QUINQUESPINOSUS SPINOLA, 1808 OF
MEDITERRANEAN TURKEY WITH AN UPDATED
LIST OF SUBFAMILY ROPHITINAE
(HYMENOPTERA: HALICTIDAE) OF TURKEY
17
TÜRKIYE’DEKI ROPHITINAE (HYMENOPTERA: HALICTIDAE) ALTFAMILYASININ GÜNCEL
LISTESI ILE TÜRKIYE’NIN AKDENIZ HAVZASINDAKI ROPHITES ALGIRUS PÉREZ, 1895 AND
ROPHITES QUINQUESPINOSUS SPINOLA, 1808 ÜZERINE NOTLAR
Fatih DİKMEN* and Ahmet Murat AYTEKİN*
Summary: The study was conducted in the Mediterranean region of southern Turkey. Two Rophitinae
(Halictidae: Hymenoptera) species, Rophites algirus Pérez, 1895 and Rophites quinquespinosus Spinola,
1808 were considered. Taxonomical identification keys, distribution maps and flower visits of these species were given. Detailed microscopic photos and drawings of some important morphological features
were also revealed. Besides, the literature on the fauna of the subfamily Rophitinae was reviewed to establish an updated species list of Turkey. As a result of this overview eight species seem possibly endemic
for Turkey.
Key Words: Halictidae, Rophitinae, Fauna, Distribution, Turkey
Özet: Çalışma Türkiye’nin güneyindeki Akdeniz Bölgesi’nde gerçekleştirildi. İki Rophitinae (Halictidae:
Hymenoptera) türü, Rophites algirus Pérez, 1895 and Rophites quinquespinosus Spinola, 1808 ele alındı.
Bu türlere ait taksonomik tanı anahtarları, yayılım haritaları ve bitki ziyaretleri belirtildi. Bazı önemli
morfolojik karakterlere ait fotoğraflar ve çizimler de ayrıca gösterildi. Bunların yanında, Türkiye’deki
güncel tür listesinin elde edilebilesi için, Rophitinae altfamilyası faunası üzerine yapılmış olan literatür
derlendi. Bu genel taslağın bir sonucu olarak, sekiz türün Türkiye için muhtemelen endemik olabilecekleri görüldü.
Anahtar Kelimeler: Halictidae, Rophitinae, Fauna, Yayılım, Türkiye
*Hacettepe University Faculty of Science, Department of Biology, 06800 Beytepe, Ankara, Turkey
Corresponding Author E-mail: [email protected]
This study based on part of the PhD thesis of F. Dikmen submitted to Hacettepe University Institute of Science in 22.06.2012.
MELLIFERA
Introduction
Halictidae (Apiformes: Apoidea: Hymenoptera) is one
of the largest families of all bees (Pesenko et al. 2000;
Michener 2007). It contains four subfamily (Rophitinae, Nomiinae, Nomioidinae, Halictinae) according
to Michener (2007) and nearly 3500 species (Pesenko
2007) in the world. From these, Rophitinae is one the
most interesting one because of their unique morphology and biological features. Rophitinae members can be
easily separated from other groups by such brief characters; antennal sockets placed on lower half of face
and clypeus shorter than labrum (Pesenko et al. 2000).
Apart from these, they exhibit solitary life and they are
mostly oligolectic unlike the other Halictidae members
(Pesenko et al. 2000). Patiny and Michez (2006) reported that Systropha spp. are the most typical ones that
show narrow plant choice and especially oligoleg on
Convovulus L. spp. (Convolvulaceae). Moreover, Patiny et al. (2007) suggested that there should be an evolutionary tendency for Rophitinae species to specialize
on the members of certain plant groups.
This subfamily contains nearly 200 species in the
world and half of them are distributed in Palaearctic
region (Pesenko et al. 2000). Generally in West Palaearctic Region and also locally in Turkey it is represented by four genera: Dufourea Lepeletier, Morawitzia Friese, Rophites Spinola, and Systropha Illiger
(Michener 2007). Among them, the most diverse genus is Dufourea and it has generally Holoarctic distribution. Whereas members of the genera Systropha,
Morawitzia and Rophites show more likely restricted
distributions (Pesenko et al. 2000; Michener 2007).
On the other hand there are many tasks had to be done
for exact evolutionary explanations. Especially there
are still no sufficient and up to date knowledge on the
distributions of Rophitinae groups for Turkey. The
leading studies which also include scrappy data on the
Rophitinae fauna of Turkey are Ebmer (1987; 1988;
1993) and Schwammberger (1976). Moreover the information about the members of the genus Rophites
Spinola of Turkey is very scanty.
For these reasons, this study aimed to make contributions on the Rophitinae fauna of Turkey. Rophites
algirus Pérez, 1895 and Rophites quinquespinosus
Spinola, 1808 were considered taxonomically. Sec-
18
ondly the scattered literature data were evaluated to
figure out the updated faunistic list of the subfamily
for Turkey.
Materials and Methods
The study was conducted at Mediterranean Region of
Southern Turkey. Field studies were performed during
the spring and summer seasons of 2008 and 2009. Random sampling protocol was used and vegetation boundaries were followed in collecting bees. Bee specimens
were collected via hand nets and aspirators. Meanwhile, the plants that have been visited by bees were
also recorded or collected for diagnosis. Captured bee
samples and collected plants were properly prepared
for collections. All of these specimens were deposited in the Department of Biology, Faculty of Science,
Hacettepe University, Ankara (Turkey). GPS coordinates were taken by Garmin Etrex H®. Materials were
examined with stereoscopic microscopes for diagnosis.
Identification of the bee specimens were made according to Warncke (1980), Ebmer and Schwammberger
(1986) and Niu et al. (2005). Identification of plant
specimens were made by Demet Töre (Department of
Biology, Faculty of Science, Hacettepe University) and
The International Plant Names Index (IPNI 2008) were
followed for the author names of the plant taxa. Taxonomical identification key for two species were prepared. Some important morphological features including the dorsal view of genitalia of these species were
also photographed. Ebmer and Schwammberger (1986)
was followed for genitalia inspection.
Ecoregion map (fig.1) was prepared with CFF 2.0 (Carto Fauna-Flora; Barbier and Rasmont 2000) and modified with Adobe Photoshop© v7.0 for a better visualization. We followed the explanation of borders of West
Palaearctic Region proposed by Patiny et al (2009).
Distribution maps (fig. 4) for the Rophites algirus and
R. quinquespinosus was also prepared with CFF 2.0
(Barbier and Rasmont 2000: Carto Fauna-Flora). Species lists for regions and subregions were prepared according to Schwammberger (1976), Ebmer (1987; 1988;
1993), Baker (1996); Pesenko (1998); Pesenko et al.
(2000), Patiny (2003; 2004), Niu et al. (2005), Patiny
and Michez (2006), Pesenko and Astafurova (2006),
Ascher et al. (2009). The species are listed in tables are
in the alphabetical order (tab.1 and tab.3).
19
Results
Species identification key for female
1. Frons with thick spines medially, with 7 – 8 spines
under median ocel and 8 – 10 spines laterally (fig. 2a)
……………………………………… R. algirus Pérez
1’. Frons with thick spines medially, with 4 – 5 spines
under median ocel and 4 – 5 spines laterally (fig. 2b)
……………………….. R. quinquespinosus Spinola
Species identification key for male
1. S6 with a slight thin process longitudinally; distal
lobe of S8 thin; gonostylus narrower and more cylindrical (fig. 2c, 2e, 3a) ………………R. algirus Pérez
1’. S6 with thick and obvious process longitudinally;
distal lobe of S8 thick; gonostylus broader and
more foliate like (fig. 2d, 2f, 3b) ……. R. quinquespinosus Spinola
Rophites algirus Pérez, 1895
Distribution: Turkey, Caucasus (Pesenko et al. 2000);
Bulgaria, Morocco, France, Iran, Italy, Tunisia,
Ukraine (Ascher et al. 2009).
Inspected Material: 18-VII-2008, 37°16’70”N,
30°07’27”E, 1963 m, Söbüce Yayla, Antalya, 1♀; 22VII-2008, 36°20’53”N, 32°35’22”E, 2225 m, Anamur
- Ermenek road Gazipaşa cross, Anamur, Mersin, 1♂;
07-VI-2009, 36°21’59”N, 33°13’63”E, 1241 m, Gülnar
- Ermenek road, 5 km to Akova, Mersin, 2♂♂; 23VII-2009, 36°26’84”N, 32°47’30”E, 1752 m, Anamur
- Ermenek road, Gazipaşa cross, Ermenek, Karaman,
1♀ (fig. 4a).
Visited Flowers: Phlomis armeniaca Willd., P. monocephala P.H.Davis, P. sieheana, Stachys byzantina C.
koch. (Lamiaceae).
Rophites quinquespinosus Spinola, 1808
Distribution: Europe, China, Western Russia, Caucasus,
Kazakhstan, Kirgizstan, Turkey (Ascher et al. 2009).
Inspected Material: 23-VII-2009, 36°26’84”N,
32°47’30”E, 1752 m, Anamur - Ermenek road
Gazipaşa cross, Ermenek, Karaman, 2♀♀ (fig. 4b).
Visited Flowers: Ballota saxatilis Sieber ex C.Presl.
(Lamiaceae).
Biogeographical results
The subfamily Rophitinae is represented by 95 species
in Palaearctic region (Pesenko and Astafurova 2006).
As a result of our literature research we can suppose
that 29 of them (13 from Dufourea; 11 from Rophites;
three from Morawitzia; and two from Systropha) are
recorded from Turkey (tab.1). These numbers also constitute nearly 30% of Palaearctic fauna and also close
with the richness of the fauna of Europe mainland
(tab.2). Moreover, eight of them are possibly endemic
for Turkey (tab.1). In addition to these data, twelve Rophitinae spp. were also listed according to their close
distribution within adjacent boundaries (tab.3).
Discussion
During our field studies in Mediterranean Region of
Turkey only two species belong to Rophites spp. were
determined. However it was interesting that all specimens were captured at high elevation (between 12002200 meters). Secondly it is important that Rophites
algirus and R. quinquespinosus are captured just only
on Lamiaceae members. This may be counted as a kind
of clue about their oligolectic feeding behavior which
was also mentioned in Pesenko et al. (2000). However
because of the low sampling size it is difficult to make
inference about the flower preferences and distributions
of the Rophites species within Mediterranean Turkey.
The biogeographical results showed that Turkey is one
of the important reserves for the members of Morawitzia, Rophites and Dufourea. On the other hand
Systropha members have Palaeatrctic and African
distribution and they are hardly represented within
Turkey (tab.2). The members of Morawitzia are generally endemic to Anatolian part of Turkey and Caucasia
(Georgia and Armenia) and do not distributed in any
other parts of the world. The endemism (nearly 20%
of Palaearctic fauna) and the species richness (nearly
60% of the Palaearctic fauna) of the genus Rophites
are also high in Turkey (Table.2). These findings generally supports that Turkey is one of the most important biogeographical zones for bees with considerably
diversified fauna. The data presented here might exhibit a good ground base for bee conservation studies
in Turkey and also would be helpful in such taxonomical studies focusing on the subfamily Rophitinae.
Acknowledgements
This study partly supported by Hacettepe University
Research Foundation Project No: 0701601016.
MELLIFERA
20
Fig. 1: Subregions of West Palaearctic Region and Asia. BA: Balkans; CA: Central Asia; CAU: Caucasia; EA: Eastern
Fig. 1: Subregions of West Palaearctic Region and Asia. BA: Balkans; CA: Central Asia;
Asia; ME: Middle East; MED: Mediterranean Basin; NA: Northern Asia; SA: Southern Asia; WP: West Palaearctic Region
(includingCAU:
BA, CAU,
MED, andEA:
ME regions).
Caucasia;
Eastern Asia; ME: Middle East; MED: Mediterranean Basin; NA:
Northern Asia; SA: Southern Asia; WP: West Palaearctic Region (including BA, CAU, MED,
and ME regions).
8
9
21
Fig. 2: a-b: Female head and the spines on frons; a: Rophites algirus and b: R. quinquespinosus; c-d: Sterna VI (S6) of
male; c: Rophites algirus and d: R. quinquespinosus; Sterna VIII (S8) of male; e: Rophites algirus and f: R. quinquespinosus.
Fig. 2: a-b: Female head and the spines on frons; a: Rophites algirus and b: R.
quinquespinosus; c-d: Sterna VI (S6) of male; c: Rophites algirus and d: R. quinquespinosus;
22
MELLIFERA
Sterna VIII (S8) of male; e: Rophites algirus and f: R. quinquespinosus.
Fig. 3: Male genitalia drawing and detailed microscopic photograph: a: Rophites algirus and b: R. quinquespinosus. gb:
gonobase; gc: gonocoxite; gs; gonostylus; pv: penis valve; vs: volsella; S8: Sterna VIII.
Fig. 3: Male genitalia drawing and detailed microscopic photograph: a: Rophites algirus and
b: R. quinquespinosus. gb: gonobase; gc: gonocoxite; gs; gonostylus; pv: penis valve; vs:
volsella; S8: Sterna VIII.
10
23
11
Fig. 4: Mediterranean distributions of a: Rophites algirus and b: R. quinquespinosus.
Fig. 4: Mediterranean distributions of a: Rophites algirus and b: R. quinquespinosus.
Table.1: The list of the Rophitinae Species of Turkey. 1: Ascher et al. (2009); 2: Ebmer
(1987); 3: Pesenko (1998); 4: Ebmer (1988); 5: Ebmer (1993); 6: Schwammberger (1976); 7:
Pesenko et al. (2000); 8: Baker (1996); Ref: References.
Species
Subgenus
Ref.
Distribution
Dufourea armenia Ebmer, 1987
Cyprirophites 1; 2
Dufourea atrata (Warncke, 1979)
Dufourea
1, 3
Endemic - Caucasia
Dufourea caelestis Ebmer, 1987
Dufourea
1; 2
Endemic
Dufourea cypria Mavromoustakis, 1952
Dufourea
1
East Mediterranean
Dufourea graeca Ebmer, 1976
Halictoides
1; 2
Caucasia, Balkan
Dufourea longicornis (Warncke, 1979)
Cyprirophites 1
Caucasia
East Mediterranean,
Middle East, West Part of
MELLIFERA
24
Table 1: The list of the Rophitinae Species of Turkey. 1: Ascher et al. (2009); 2: Ebmer (1987); 3: Pesenko
(1998); 4: Ebmer (1988); 5: Ebmer (1993); 6: Schwammberger (1976); 7: Pesenko et al. (2000); 8: Baker
(1996); Ref: References.
Species
Subgenus
Ref.
Distribution
Dufourea armenia Ebmer, 1987
Cyprirophites
1; 2
Caucasia
Dufourea atrata (Warncke, 1979)
Dufourea
1, 3
Endemic - Caucasia
Dufourea caelestis Ebmer, 1987
Dufourea
1; 2
Endemic
Dufourea cypria Mavromoustakis, 1952
Dufourea
1
East Mediterranean
Dufourea graeca Ebmer, 1976
Halictoides
1; 2
Caucasia, Balkans
Dufourea longicornis (Warncke, 1979)
Cyprirophites
1
East Mediterranean, Middle East, West
Part of East Asia
Dufourea pontica (Warncke, 1979)
Halictoides
1; 2
Caucasia
Dufourea quadridentata (Warncke, 1979)
Dufourea
1
Endemic
Dufourea salviae Ebmer, 2008
Cyprirophites
1
Endemic
Dufourea schmiedeknechtii (Kohl, 1905)
Halictoides
1; 2; 3
North Asia; Caucasia; Europe
Dufourea wolfi Ebmer, 1989
Dufourea
1
Balkans
Morawitzia fuscescens Friese, 1902
Morawitzia
1
Endemic
Morawitzia mandibularis Alfken, 1935
Morawitzia
1
Caucasia
Morawitzia panurgoides Friese, 1902
Morawitzia
1
Caucasia
Rophites algirus Pérez, 1895
Rophites
1; 4
West Palaearctic
Rophites anatolicus (Schwammberger, 1975)
Rophitoides
1
Endemic
Rophites canus Eversmann, 1852
Rophitoides
1; 4
Trans-Palaearctic
Rophites caucasicus Morawitz, 1875
Rophites
1; 5
Caucasia
Rophites clypealis Schwammberger, 1976
Rophites
1; 6
Pontic
Rophites foveolatus Friese, 1900
Rophites
1; 5
Caucasia, South Europe
Rophites gusenleitneri Schwammberger, 1971
Rophites
1; 5
Endemic
Rophites hartmanni Friese, 1902
Rophites
1; 4
East Europe, East Mediterranean
Rophites heinrichi Schwammberger, 1976
Rophites
1, 6
Endemic
Rophites leclercqi Schwammberger, 1971
Rophites
1, 7
Balkans
Rophites nigripes Friese, 1902
Rophites
1; 5
East Mediterranean
Rophites quinquespinosus Spinola, 1808
Rophites
1
WP, Middle East
Rophites transitorius Ebmer, 1993
Rophites
1; 5
Endemic
Systropha curvicornis (Scopoli, 1770)
Systropha
1; 4
West Palaearctic and East Asia
Systropha planidens Giraud, 1861
Systropha
1; 4; 8
Europe and Middle East
25
Table 2: Comparison of the Rophitinae fauna of Palaearctic Region, Europe and Turkey.
Genus
Palaearctic
Turkey
Europe
(Number of Species)
(Number of Species)
(Number of Species)
General Distribution
Dufourea
Holarctic
54
11 (3 endemic)
17
Rophites
Palaearctic
21
13 (4 endemic)
10
17
2 (no endemism)
2
3
3 (1 endemic)
-
Mainly Palaearctic
Systropha
and Africa
Morawitzia
Caucasia
Table 3: The list of the Rophitinae species of adjacent boundaries. 1: Ascher et al. (2009); 2: Ebmer (1987).
Species
Reference
Distribution
Dufourea alpina Morawitz, 1865
1
Mediterranean (South Europe)
Dufourea bytinskii Ebmer, 1999
1
East Mediterranean
Dufourea dentiventris (Nylander, 1848)
1
Europe, East Asia
Dufourea goeleti Ebmer, 1999
1
East Mediterranean
Dufourea halictula (Nylander, 1852)
1
Europe, Caucasia
Dufourea inermis (Nylander, 1848)
1
Europe, Northeastern and East Asia
Dufourea iris Ebmer, 1987
1; 2
Balkans
Dufourea minuta Lepeletier, 1841
1
Europe, East Asia
Dufourea paradoxa (Morawitz, 1867)
1
West Europe, Central Asia, East Asia
Dufourea similis Friese, 1898
1
North Africa, East Mediterranean
Dufourea trigonellae Ebmer, 1999
1
East Mediterranean
Rophites schoenitzeri Dubitzky, 2005
1
Caucasia
MELLIFERA
References
Ascher J.S., Rozen Jr. J.G. and Schuh T. 2009. Discoverlife
website. Apoidea species guide. http://www.discoverlife.org/mp/20q?guide=Apoidea_species
Barbier Y. and Rasmont P. 2000. Carto Fauna-Flora 2.0.
Guide d’utilisation. Université de UMH-Hainaut,
UMH, Belgique, pp. 59.
Baker D.B. 1996. Notes on some palaearctic and oriental
Systropha, with descriptions of new species and a
key to the species (Hymenoptera: Apoidea: Halictidae). Journal of Natural History, 30: 1527-1547.
Ebmer A.W. 1987. Die westpaläarktischen Arten der Gattung Dufourea Lepeletier 1841 mit illustrierten
Bestimmungstabellen. Linzer Biologische Beiträge, 19: 43-56.
Ebmer A.W. 1988. Kritische liste der nicht-parasitischen
Halictidae Österreichs mit Berücksichtigung aller
mitteleuropäischen Arten (Insecta: Hymenoptera:
Apoidea: Halictidae). Linzer biol. Beitr, 20 (2):
527-711.
Ebmer A.W. 1993. Die Bienengattung Rophites Spinola 1808
– Erster Nachtrag. Linzer Biologische Beiträge,
25: 3-14.
Ebmer A.W. and Schwammberger K.H. 1986. Die Bienengattung Rophites Spinola 1808 (Insecta: Hymenoptera: Apoidea: Halictidae: Dufoureinae). Illustrierte Bestimmungstabellen. Senckenbergiana
biol., 66: 271-304.
26
Annales de la Société entomologique de France,
42(1):27-44.
Patiny S., Michez D. and Danforth B.N. 2007. Phylogenetic
relationships and host-plant association within the
basal clade of Halictidae (Hymenoptera, Apoidea).
Cladistics, 23:1-15.
Patiny S., Rasmont P. and Michez D. 2009. A survey and
review of the status of wild bees in the West Palaearctic region. Apidologie, 40 (2009): 313-331.
Pesenko Yu.A. 1998. New and little known bees of the genus
Dufourea Lepeletier (Hymenoptera, Halictidae)
from the Palaearctic Region. - Ent. Obozrenie (St.
Petersburg), 77 (3): 670-686 [in Russian with English summary. English translation in Ent. Review,
78 (5): 598-612].
Pesenko Yu.A. 2007. The family Halictidae (Hymenoptera):
General. In: A key to insects of the Russian Far
East. Vol. IV, pt 5. Vladivostok (Dal’nauka): 745754. [in Russian]
Pesenko Yu.A., Banaszak J., Radchenko V.G. and Cierzniak
T. 2000. Bees of the Family Halictidae (Excluding Sphecodes) of Poland: Taxonomy, Ecology,
Bionomics. Bydgoszcz, Poland. Bydgoszcz Press.
p. 348.
Pesenko Yu.A. and Astafurova Yu.V. 2006. Contributions to
the halictid fauna of the Eastern Palaearctic Region: subfamily Rophitinae (Hymenoptera: Halictidae). Entomofauna, 27(27): 317–356.
IPNI 2008. The International Plant Names Index, www.ipni.
org.
Schwammberger K.H. 1976. Zwei neue Rophites-Arten aus
der Türkei. Ent. Ztschr. 86: 225-228.
Michener C.D. 2007. The Bees of the World. 2nd edition.
John Hopkins Univ. Press, Balitimor, USA. 953 p.
Warncke K. 1980. Rophites quinquespinosus Spinola und
R. trispinosus Pérez eine oder zwei Bienenarten?
(Apidae, Halictinae). Entomofauna, 1/3: 37-52.
Niu Z., Wu Y. and Huang D. 2005. A taxonomic study of
the four genera of the subfamily Rophitinae from
China (Hymenoptera: halictidae). The Raffles
Bulletin of Zoology. 53: 47-58.
Patiny S. 2003. Revision of the subgenus Dufourea (Flavodufourea) Ebmer, 1984 (Hymenoptera, Halictidae, Rophitinae) and description of a new species
D.(Flavodufourea) ulkenkalkana sp.nov. from Kazakhstan. Zootaxa, 255: 1-8.
Patiny S. 2004. Description of two new Systropha Illiger
1806 (Hymenoptera, Halictidae, Rophitinae).
Linzer Biologische Beiträge. 36 (2): 907-912.
Patiny S. and Michez D. 2006. Phylogenetic analysis of the
Systropha Illiger, 1806 (Hymenoptera: Apoidea:
Halictidae) and description of a new subgenus.
MELLIFERA
12-24:27-32 (2012) HARUM
RESEARCH ARTICLE
27
CHEMICAL COMPOSITION OF PROPOLIS SAMPLES
COLLECTED FROM TEKIRDAG-TURKEY
TEKİRDAĞ-TÜRKİYE’DEN TOPLANAN PROPOLİS ÖRNEKLERİNİN KİMYASAL İÇERİĞİ
Ömür Gençay Çelemli *, Kadriye Sorkun*, Bekir Salih**
Summary: The aim of this study is to investigate the chemical compositions of propolis samples which were collected from Tekirdağ city of Turkey. A total of 92 different propolis samples
collected from eight towns of Tekirdağ were examined by GC-MS (Gas Chromatography and
Mass Spectrometry) to determine chemical composition and establish the chemical profile of
Tekirdağ propolis.
According to the GC-MS results, the compound; aldehydes, alcohols, aliphatic acids and their
esters, flavonoids, hydrocarbons, carboxylic acid and their esters, cinnamic acids and their
esters ketones, were determined in various amounts. Among these compounds the flavonoids
were found in all samples and in higher amounts compare to the other compounds.
Keywords: propolis, GC-MS, Tekirdağ, chemical profile, flavonoid
Özet: Bu çalışmanın amacı Türkiye’nin Tekirdağ ilinden toplanan propolis örneklerinin kimyasal içeriğini araştırmaktır. Tekirdağ’ın sekiz ilçesinden toplanan toplamda 92 örneğin kimyasal içeriği GC-MS ile (Gaz Kromatografisi ve Kütle Spektrometresi) incelenmiş ve Tekirdağ
propolisinin kimyasal profili oluşturulmuştur.
GC-MS sonuçlarına göre aldehidler, alkoller, alifatik asit ve esterleri, flavonoidler, hidrokarbonlar, karboksilik asit ve esterleri, sinamik asit ve esterleri, keton bileşikleri değişik miktarlarda saptanmıştır. Bu bileşikler arasında flavonoidler tüm örneklerde belirlenmiş ve diğer
bileşiklere göre daha yüksek oranlarda olduğu bulunmuştur.
Anahtar kelimeler: propolis, GC-MS, Tekirdağ, kimyasal profil, flavonoid
*Hacettepe University Faculty of Science, Department of Biology, 06800 Beytepe, Ankara, Turkey
**Hacettepe University Faculty of Science, Department of Chemistry, 06800 Beytepe, Ankara, Turkey
Corresponding Author E-mail: [email protected]
This study based on part of the PhD thesis of Ö. G. Çelemli.
MELLIFERA
Introduction
Propolis or bee glue is a sticky dark-colored material
that honey bees collect from plants and use it in the
hive: they apply it to seal the walls, to strengthen the
borders of combs, to line all cells inside, to embalm
dead invaders (Bankova 2005). It is also well known
that the propolis possesses antibacterial, antifungal
and antiviral properties and many beneficial biological activities such as antiinflammatory, antiulcer,
local anesthetic, hepatoprotective, antitumor, immunostimulating etc (Bankova et al. 2000). Bees use it,
therefore, as a protective barrier against their enemies
(Burdock 1998). However, propolis is being used in
the traditional medicine since 3000 BC, in Egypt.
For propolis production, bees use natural materials
resulting from a variety of botanical processes in different parts of plants. These are substances actively
secreted by plants as well as substances exuded from
wounds in plants: lipophilic materials on leaves and
leaf buds, gums, resins, lattices etc. The plant origin
of propolis determines its chemical diversity. Bee
glue’s chemical composition depends on the specify
of the local flora at the site of collection and thus on
the geographic and climatic characteristics of this site
(Bankova 2005).
Propolis generally contains 50% resin, 30% wax, 5%
pollen, 10% aromatic oils, and 5% other organic residues. Literature reported some important biological
activities of propolis. The biological activities were
verified due to the content of flavonoids, aromatic acids and esters present in the propolis (Lee et al. 2007).
Material and Methods
Propolis samples
In 2007-2008 the propolis samples were collected from
the hives of Tekirdağ. The hives from eight towns
(Çerkezköy, Çorlu, Hayrabolu, Malkara, Merkez,
Muratlı, Saray, Şarköy) of Tekirdağ choosed according to the sampling method. By this method 92 bee
farms were choosen to collect propolis. So the study
carried on with 92 propolis samples. The number of
beehives choosen by sampling method is given in Table 1. Propolis samples were collected from the edges
of frames by scraping with a spatula.
28
Extraction and sample preparation
Each frozen propolis sample was grained and dissolved in ethanol (96%) with a ratio of 1/3. Then ,the
mixture kept in tightly closed bottle and in an incubator at 30°C for two weeks. After incubation period,
the supernatant was filtered twice with Watman No 4
and No 1 filter papers. The final solution, (1:10, w/v),
called Ethanol Extracts of propolis (EEP) was evaporated until completely dryness. About 5 mg of dry
substance were mixed with 75 µl of dry pyridine and
50 µl bis (trimethylsilyl) trifluoroacetamide (BSTFA),
heated at 80°C for 20 min and the final supernatant
was analyzed by GC-MS (Gençay and Salih 2009).
GC-MS analysis
A GC 6890N from Agilent (Palo Alto, CA, USA) coupled with mass detector (MS5973, Agilent) was used
for the analysis of EEP samples. Experimental conditions of GC-MS system was as follows: DB 5MS
column (30 mx 0.25mm and 0.25 µm of film thickness) was used and flow rate of mobile phase (He) was
set at 0.7 ml/min. In the gas chromatography part,
temperature was kept for 1 min at 50 °C and then increased to 150 °C with 10 °C/min heating ramp. After
this period, temperature was kept at 150 °C for 2 min.
Finally, temperature was increased to 280 with 20 °C/
min. heating ramp and kept at 280 °C for 30 min.
Organic compounds in propolis samples were identified using standard Willey and Nist Libraries available in the data acquisiton system of GC-MS, if the
comparison scores were obtained higher than 95%.
Otherwise, fragmentation peaks of the compounds
were evaluated and the compounds were identified using our memorial background for the identification of
the compounds appeared in GC-MS chromatograms.
For the quantification of the compounds in the ethanol
extract, no internal and external standards were used;
only percentage reports of the compounds in the sample were used. This was the standard way to quantify
most organic compounds in the propolis sample, thus
reducing the relative error in <5% .
Results and Discussion
According to the GC-MS results aldehydes, alcohols,
aliphatic acids and their esters, flavonoids, hydrocarbons, carboxylic acid and their esters, ketones, cin-
29
namic acids and their esters were determined in various amounts in the investigated 92 samples. Among
these compounds flavonoids were the compounds observed at higher ratios.
Naphthalene, 1, 2, 3 ,4 ,4a ,5 ,6 , 8a-octahydro-7-methyl-4-methylen-1-(1-methylethyl) (1.alpha., 4a.beta.,
8a.alpha.) compounds were observed frequently but in
minor amounts.
The flavonoids; “2-Propen-1-one,1-(2,6-dihydroxy4-metoxyphenyl)-3-phenyl (Pinostrobin, chalcone),
4, 5-Dihydroxy-7-methoxyflavanone, 4H-1-benzopyran-4-one, 5-hydroxy-2-(4-hydroxyphenyl)-7methoxy-(Tetrochrysin), 4H-1-Benzopyran-4-one,2,3-dihydroxy-5,7- dihydroxy-2-phenyl (Pinocembrin), 4H1-Benzopyran-4-one,5-hidroxy-7-methoxy-2-phenyl,
4H-1-Benzopyran-4-one,3,5,7-trihydroxy-2-phenyl
(Galangin), 4H-1-Benzopyran-4-one 5,7 dihydroxy2-phenyl (Chrysin), 4H-1-Benzopyran-4-one,5,7dihydroxy-2-(4-hidroxyphenyl) (Acacetin)” were observed in Tekirdağ samples.
As carboxylic acids and their esters ; “Heptadecanoic
acid,15-mehyl-ethyl ester, 4-Pentenoic acid, 5-phenyl,
Benzoic acid, 4-pentenoic acid, 5-phenyl-cyclopropancarboxylic acid, 2-phenyl-, methyl ester, 2-butenoic
acid, 2-methyl, pentadecanoic acid ethyl ester, 1,2-benzenedicarboxylic acid diis ooctyl ester” were found to
be main compounds.
The compound belong to the aldehydes group were
observed in minor amounts.
From aliphatic acids and their esters group mostly observed compounds were; Ethyl oleat, Hexadecanoic acid
ethyl ester (palmitic acid ethyl ester), Octadecanoic acid
ethyl ester (stearic acid ethyl ester), 9-Octadecanoic acid,
Linoleic acid ethyl ester, 2-Butenoic acid, 2 methyl, Hexadecanoic acid, 9, 12-Octadecadienoic acid ethyl ester,
Decanoic acid ethyl ester, Dodecanoic acid ethyl ester,
Heptadecanoic acid, 15-methyl-,ethyl ester.
From hydrocarbons group; 1,19-Eicosadiene, 10 –
heneicosene (c,t), 17-Pentatriacontene, 1-Docosane,
1-Heptadecane, 1-hexacosane, 1,13-Tetradecadiene,
1-Nonadecane 3-Eicosane, 7-Pentadecine, 9–Tricosane, (Z)-, 9,10–Antracenedione, 1-hidroxi–2–
(hidroximethyl), 9,10-Anthracenedione, 1-hidroxy2-(hydroxymethyl)-, 9-Hexacosane, 9-Nonadecene
Nonadecane, Octadecane, Pentacosane, Benzofuran, 2,3-dihydro-, Siclotetracosane, Docosane,
Eicosane, Tricosene, Heneicosane, Heptacosene,
Z-12-pentacosene, Z-5-Nonadecane, Siclotriacontane, 1,3,5,7-Cyclooctatetracane, Naphthalene,
1,2,3,5,6,7,8,8a –octahydro–1, 8a–dimethyl–7–( 1 –
methylethenyl)-, [1R–(1.alpha, 7. beta, 8a alpha.)]-,
Naphthalene, 1,2,3,4-tetrahydro-1,6-dimethyl-4-(1methylethyl)-, (1S-cis)-, Naphthalene,1,2,3,5,6,8ahexahydro-4,7-dimethyl-1-(1-methylethyl)-,(1S-cis),
From cinnamic acid and its esters group; “benzyl cinnamate, 2-Propenoic acid ,3-phenyl- (cinnamic acid),
Benzenepropanoic acid (Hydrocinnamic acid), Benzenpropanoic acid methyl ester, 2-propenoic acid,
3-phenyl-, methyl ester, 3,4-dimethoxycinnamic acid,
3-hidroxy-4-methoxycinnamic acid, Benzyl benzoate“ were observed in minor ratios.
Discussion
Among the determined compounds, the flavonoids
were observed in quite higher amounts. In the most
of the samples, chrysin a kind of flavonoid was determined. It has anti-tumor (Hladon et al., 1980), antiHelicobacter pylori (Itoh et al., 1994), anti-inflammatory (Shin et al., 2009), anti-oxidant, antiviral, antidiyabetogenic, anti-axyoletic (Zheng et al., 2003) effects. So we can say that the 86 samples from Tekirdağ
region, contain chrysin, may have valuable biological
effects for further studies.
Another kind of flavonoid; pinocembrin was determined in 86 Tekirdağ samples. This compound
has bacterioteriostatic (Amoros et al., 1992), antimould (Miyakado, 1976), antimicrobial, anti-micotic
(Metzner et al., 1979), anti-oxidant, anti-inflammatory (Gao et al.,2008) and local anesthesic effects. Also
its anti microbial effect to Alternalis fungus was determined (Miyakado et al., 1976).
Galangin is an other flavonoid found in 53 Tekirdağ
samples. It has bacteriostatic (Amoros et al., 1992;
Pepeljnjak, 1982), antimicrobial, anti-micotic
(Metzner et al., 1979), anti-Helicobacter pylori (Itoh
et al., 1994) activities.
MELLIFERA
Comparing with previous studies in flavonoid contents base, similiar to our results galangin (in Bursa, Muğla, İzmir, Ankara samples) (Velikova et al.,
2000), chrysin; (in Bursa, İzmir, Ankara, Elazığ and
Erzincan, Erzurum samples) (Velikova et al., 2000;
Kılıç et al. 2005; Silici and Kutluca 2005; Seven et
al. 2005), pinocembrin (in Bursa, İzmir, Muğla, Ankara samples), 4,5-Dihidroksi-7-methoxyflavanon (in
Ankara and Erzincan samples) (Kılıç et al., 2005) ,
pinostrobin chalcone ( in Erzincan sample) (Gençay,
2004), acacetin (in Erzurum sample) (Silici ve Kutluca 2005) had been observed by other researchers.
Benzaldehyde, 3-hydroxy-4-methoxy was found in
Ankara and Erzincan samples by Kılıç et al. is similiar to our results (2005).
According to the GC-MS results, “ octadecanoic acid”
from the frequently observed compounds belong to the
aliphatic acids and their esters were found in Ankara
and Bursa samples in previous studies. From identified
aliphatic acids and esters in Tekirdağ samples , “ethyl
oleate” was found in Ankara, Kemaliye-Erzincan
samples by Kılıç et al. (2005), “hexadecanoic acid“ in
Ankara, Bursa, İzmir samples by Velikova et al (2000).
Benzoic acid, the mostly observed compound from
carboxylic acid and esters group in Tekirdağ samples,
was found in the samples collected from Ankara,
Bursa, İzmir, Muğla (Velikova et al., 2000), and Zonguldak samples (Girgin et al., 2009) by other researchers. This compound was identified in 77 Tekirdağ
samples in our study and its bacteriostatic, antiseptic
properties were showed by former researchs.
Cinnamic acid, that is important for biological activities of propolis, was observed frequently (in 35
Tekirdağ samples) but in minor amounts in the in-
30
vestigated samples. This compound also has been observed in Ankara, İzmir, Muğla samples by Velikova
et al.(2000).
The determined amounts of flavonoid, hydrocarbon,
carboxylic acids and their esters were compared by
ANOVA-Duncan analysis in town base ( Table 2-4).
According to the results of Anova-Duncan analysis,
the amount of flavonoid in the propolis samples belong to the eight towns are very similiar and divided
only two groups. As seen in table 2 values are very
similiar to each other. Malkara samples contain the
highest flavonoid ratios, Çerkezköy samples contain
lower flavonoid content compare to the other towns.
It is noticed from table 3 that, there is a big difference
between Merkez and Şarköy samples according to the
hydrocarbon contents.
According to the results there is a big difference in
the amount of carboxylic acids and esters between
Şarköy and Saray samples (Table 4).
As a result Tekirdağ propolis samples are valuable for
further studies. All the samples are rich in flavonoid
content and especially Malkara samples has the highest flavonoid content that can give antimicrobial, antioxidant, etc. activities to propolis samples. Also the
samples may have anti viral, anti micotic, anti inflammatory, anti Staphylococcus aureus activities owing
to the cinnamic acids and esters contents.
Acknowledgement
This research is supported by Hacettepe University
Scientific Research and Development Office(Project
Number: 0701601008).
31
Table 1. The number of collected propolis samples and collecting areas
Number
Towns
The number
beehives (Nh)
1
Çerkezköy
2
Çorlu
3
4
The number of samples
that must be collect (nh)
The number of collected
samples
35
7
7
44
8
8
Ereğli
12
2
-
Hayrabolu
48
9
9
5
Malkara
74
14
14
6
Merkez
164
31
31
7
Muratlı
45
9
9
8
Saray
65
12
12
9
Şarköy
11
2
2
9 TOWNS
497 BEEHIVES
94
92
TOTAL
of
registered
Table 2. The statistical comparing of eight towns of Tekirdağ city according to the Flavonoids
Table 2. The statistical comparing of eight towns of Tekirdağ city according to the Flavonoids
Table 2. amount
The statistical comparing of eight towns of Tekirdağ city according to the Flavonoids amount
amount
1; Çerkezköy
samples,
Çorlu
samples,3;
3; Hayrabolu,
Hayrabolu,
4;
5;5;Merkez,
Muratl,
7;
8; Şarköy
*1; Çerkezköy samples,
2; Çorlu
samples,
3; Hayrabolu,
4; Malkara,
5; Merkez,
6; 6;
Muratlı,
7;Saray,
Saray,
8;8; Şarköy
*1; *Çerkezköy
samples,
2;2;Çorlu
samples,
4;Malkara,
Malkara,
Merkez,
6;
Muratl,
7;
Saray,
Şarköy
Table
Thestatistical
statisticalcomparing
comparing of
of eight
eight towns
to to
the the
Table
3. 3. The
towns ofof Tekirdağ
Tekirdağcity
cityaccording
according
hydrocarbons
amount
hydrocarbons
amount
Table 3. The statistical comparing of eight towns of Tekirdağ city according to the hydrocarbons amount
*1; Çerkezköy samples,
2; Çorlusamples,
samples,
3; Hayrabolu,
4; Malkara,
5; Merkez,
6; 6;
Muratlı,
7;Saray,
Saray,
8; Şarköy
*1; Çerkezköy
2; Çorlu
samples, 3; Hayrabolu,
4; Malkara,
5; Merkez,
Muratl, 7;
8; Şarköy
*1; Çerkezköy samples, 2; Çorlu samples, 3; Hayrabolu, 4; Malkara, 5; Merkez, 6; Muratl, 7; Saray, 8; Şarköy
MELLIFERA
32
Table 4. The statistical comparing of eight towns of Tekirdağ city according to the carboxylic
acids and their esters amounts
Table 4. The statistical comparing of eight towns of Tekirdağ city according to the carboxylic acids and their
esters amounts
Çerkezköy
2; Çorlu samples,
3; Hayrabolu,
4; Malkara,
5; Merkez,
Muratl,7;7;Saray,
Saray, 8;
*1; Çerkezköy*1;
samples,
2; samples,
Çorlu samples,
3; Hayrabolu,
4; Malkara,
5; Merkez,
6; 6;
Muratlı,
8; Şarköy
Şarköy
References
Amoros M., Sauvager F., Girre L., Cormier M.1992. In vitro
antiviral activity of propolis. Apidologie. 231-240.
Bankova V. 2005. Recent trends and important developments
in propolis research. eCAM. 2(1): 29Bankova V.-S.,
De Castro S.-L and Marcucci M.-C.2000. Propolis:
recent advances in chemistry and plant origin. Apidologie. 31: 3-15.
Burdock G- A. 1998. Review of the biological properties and
toxicity of bee propolis (propolis). Food Chemistry
and Toxicology.36, 347–363.
Gao M., Zhang W., Liu Q., Hu J., Liu G., Du G.2008. Pinocembrin prevents glutamate-induced apoptosis in SHSY5Y neuronal cells via decrease of bax/bcl-2 ratio,
Eur.J. Pharmacol. 591: 73-79.
Gençay Ö. Kemaliye-Erzincan Yöresine Ait Propolislerin Orijini ve Kimyasal İçeriğinin Belirlenmesi. Msc thesis.
Hacettepe University, Ankara, 2004.
Gençay Ö., Salih B. 2009. GC-MS Analysis of Propolis Samples
From 17 Different Regions of Turkey, Four Different
Regions of Brazil and One From Japan . Mellifera
,9:17,19-28.
Girgin G., Baydar T., Ledochowski M., Schennach H., Bölükbaşı
D., Sorkun K., Salih B., Şahin G., Fuchs D. 2009. Immunomodulatory effects of Turkish propolis: Changes in neopretin release and tryptophan degradation.
Immunobiology. 214,129-134.
Hladon B., Bylka W., Wojtaszek ME., Skrzypczale L., Szafarele P., Chodera A., Kowalewski Z.1980. ın vitro
studies on the cytostatic activity of propolis extracts.
Arzneim.-Forsch.Drug. Res. 30; 1847-1848.
Itoh J., Chong F., Wang H., Park Y., Ikekagi M., Kilgore N.,
Lee K.. 2001. Anti-HIV activity of moronic acid derivatives and the new melliferone related triterpenoid
isolated from Brazilian propolis. Journal of Natural
products. 64 (10); 1278-1281.
Kılıç A., Baysallar M., Beşirbellioğlu B., Salih B., Sorkun K.,
Tanyüksel M. 2005. In vitro antimicrobial activity of
propolis against methicillin-resistant Staphylococcus aureus and vancomycin-resistant Enterococcus
faecium. Annals of Microbiology.55:2, 113-117.
Lee YN., Chen CR, Yang HL., Lin CC., Chang CMJ. 2007.
Isolation and purification of 3,5-diprenyl-4-hydroxycinnamic acid (artepillin C) in Brazillian propolis by
supercritical fluid extractions. Separation and Purification Technology. 54, 130-138.
Metzner J., Bekemeier H., Paintz M., Schneidewind E.1979. Antimicrobial activity of propolis and its constituents.
Pharmazie. 34(2), 97-102.
Miyakado M., Karto T., Ohno N., Mabry T.1976. Pinocembrin
and +-β-eudesmol from Hymenoclea monogyra and
Baccharis glutinosa. Phytochemisrty. 15 (5). 846.
Pepeljnjak S.1982. Growth inhibition of Bacillus subtilis and
composition of various propolis extracts. Pharmazie.
37(12), 1995-2018.
Seven TP., Seven I., Yılmaz M., Şimşek ÜG.2008. The effects of
Turkish propolis on growth and carcass characteristics in broiler under heat stress. Animal feed Science
and technology. 146, 1-2:137-148.
Shin EK., Kwon HS., Kim YH., Shin HK., Kim JK.2009. chrysin a naturel flavones, improves murine inflammatory boel diseases. Biochemical and Biophysical Research Communications. 381,502-507.
Silici S., Kutluca S. 2005. Chemical composition and antibacterial activitu of propolis collected by three different
races of honeybees in the same region. Journal of
Ethnopharmacology, 99: 69-73.
Velikova M., Bankova V., Sorkun K., Houcine S., Tsvetkova I.
And Kujumgiev A. (2000), Propolis from the Mediterranean region: chemical composition and antimicrobial activity. Z. Naturforsch C. 55, 790-793.
Zheng J., Li Y., Zhao J., Xue X., Wu L., Chen F. 2008. Geographical traceability of propolis by 13
high-performance liquid chromatography fingerprints. Food
Chemistry. 108:749-759.