Post on 28-Mar-2023
1 23
Plant Systematics and Evolution ISSN 0378-2697Volume 300Number 4 Plant Syst Evol (2014) 300:689-708DOI 10.1007/s00606-013-0913-8
Comparative pollen morphology of annualTrigonella L. (Fabaceae) in Turkey
N. Munevver Pinar, Hasan Akan, TalipCeter, Zeki Aytac, Murat Ekici, AydanAcar & Selin Akdogan
1 23
Your article is protected by copyright and
all rights are held exclusively by Springer-
Verlag Wien. This e-offprint is for personal
use only and shall not be self-archived
in electronic repositories. If you wish to
self-archive your article, please use the
accepted manuscript version for posting on
your own website. You may further deposit
the accepted manuscript version in any
repository, provided it is only made publicly
available 12 months after official publication
or later and provided acknowledgement is
given to the original source of publication
and a link is inserted to the published article
on Springer's website. The link must be
accompanied by the following text: "The final
publication is available at link.springer.com”.
ORIGINAL ARTICLE
Comparative pollen morphology of annual Trigonella L.(Fabaceae) in Turkey
N. Munevver Pinar • Hasan Akan • Talip Ceter •
Zeki Aytac • Murat Ekici • Aydan Acar •
Selin Akdogan
Received: 31 July 2013 / Accepted: 30 August 2013 / Published online: 11 September 2013
� Springer-Verlag Wien 2013
Abstract Pollen morphology of 38 Trigonella species
was investigated with light microscopies and scanning
electron microscopies. Pollen slides were prepared using
Wodehouse technique. The pollen grains of Trigonella
members are radially symmetrical and isopolar, their out-
lines are oblong in equatorial view and circular in polar
view. Amb is semicircular. The pollen grains are prolate-
spheroidal, oblate-spheroidal, suboblate, spheroidal, and
prolate with the polar axes 13.5–49.9 lm and the equato-
rial axes 10.4–43.7 lm. The smallest dimensions are
observed in T. spinosa and T. brachycarpa, and the largest
in T. carica and T. rhytidocarpa. The pollen grains of
Trigonella taxa are usually tricolporate or rarely tricolpate,
pantocolporate, tetracolporate, and tricolpodiporate. Vari-
ous ornamentation types were observed: Microreticulate
(section Samaroideae, Pectinatae, Falcatulae, Cylindricae,
Bucerates, Reflexae, Isthmocarpae, Capitatae and Foenum-
graecum), rugulate-scabrate (section Lunatae, Uncinatae),
perforate (T. halophila, T. rigida and T. carica from sec-
tion Bucerates) and retipilatae (section Biebersteinianae) at
equator region and psilate-perforate (section Samaroideae,
Pectinatae, Bucerates, Reflexae, Isthmocarpae, Foenum-
graecum, T. coelesyriaca, T. cilicica), rugulate-scabrate
(section Lunatae, Uncinatae), microreticulate (Falcatulae,
Cylindricae and Foenum-graecum) and retipilatae (section
Biebersteinianae) at polar region. Ornamentations, pollen
shape and the aperture type have been observed as
important morphological characters.
Keywords Fabaceae � Trifoliae � Trigonella �Pollen morphology � SEM � Turkey
Introduction
The tribe Trifolieae in the family Fabaceae includes six
genera: Trigonella L., Medicago Inc., Trifolium L., Meli-
lotus L., Parochetus Buch.-Ham and Ononis L. (Heyn
1981; Lock and Simpson 1991; Mabberley 1997). Trigo-
nella is one of the largest genera in the tribe Trifolieae and
comprises about 135 species widely distributed in dry
regions around the East Mediterranean, West Asia, South
Europe, North and South Africa, with only one species
growing in South Australia (Townsend and Guest 1974;
Polhill and Raven 1981, Kawashty et al. 1998; Ceter et al.
2012; Ranjbar et al. 2012a, b). It has been postulated by
Vavilov that the near Eastern region extends from Palestine
through Syria, southern Turkey into Iran and Iraq, and the
Mediterranean region including Spain, Morocco and Iran,
and that Afghanistan is the center of origin for Trigonella
(Ranjbar et al. 2012a, b). In Turkey, the genus includes 49
species with two subspecies in 13 sections growing in
different phytogeographical regions. There are 21 taxa,
with 42 % endemic to the country (Davis 1970; Akan et al.
2005; Ceter et al. 2012).
N. M. Pinar � A. Acar � S. Akdogan
Department of Biology, Science Faculty, Ankara University,
Ankara, Turkey
H. Akan
Department of Biology, Arts and Science Faculty, Harran
University, Sanlıurfa, Turkey
T. Ceter (&)
Department of Biology, Arts and Science Faculty, Kastamonu
University, Kastamonu, Turkey
e-mail: talipceter@gmail.com
Z. Aytac � M. Ekici
Department of Biology, Science Faculty, Gazi University,
Ankara, Turkey
123
Plant Syst Evol (2014) 300:689–708
DOI 10.1007/s00606-013-0913-8
Author's personal copy
Trigonella species are annual or perennial herbs with
pinnately trifoliate leaves, often emitting an odor, and like
other grain legumes, are important in food and medicine
production (Chopra et al. 1956; Girardon et al. 1986; Ba-
lodi and Rao 1991; Bhatti et al. 1996; Dangi et al. 2004).
But all the members in Turkey are annual (Davis 1970;
Akan et al. 2005).
Taxonomic problems of this genus cannot be ignored.
Species of Trigonella are similar to each other in their
morphological characters such as floral architecture and
leaf shape. Several investigators have attempted to employ
the taxonomy of the genus Trigonella. Sirjaev (1934) gave
an elaborate and systematic account of its taxonomy in
Latin. Vasil’chenko (1953a, b) published a synopsis in
Russian discussing the genus within the family Fabaceae.
The genus was divided into three subgenera (Trigonella,
Trifoliastrum and Foenum-graecum) on the ban’s of the
form and shape of calyx and pod by Hutchinson (1964) and
Tutin and Heywood (1964). Numerical taxonomic studies
were done on 62 species of Trigonella and were separated
into 12 sections (Falcatulae, Callicerates, Uncinatae, Cy-
lindricae, Samaroideae, Capitatae, Pectinatae, Erosae,
Verae, Spinosae and Foenum-graecum) of the classical
monographer (Small 1987). According to ‘‘Flora of Tur-
key’’ the genus represented with 52 taxa, after completed
this study, the number of taxa decreased to 50 (49 species
and one subspecies) in Turkey. In Turkish Flora, the genus
was revised by Davis (1970) and Akan et al. (2005) who
divided it into 13 sections: section Samaroideae, Pectina-
tae, Lunatae, Falcatulae, Verae, Cylindricae, Bucerates,
Reflexae, Isthmocarpae, Uncinatae, Capitatae, Bieberstei-
nianae and Foenum-graecum.
Generic circumscription has long been a problematic
question in the tribe Trifolieae subtribe Trigonellinae. In
particular, many studies have been undertaken on ascer-
taining the prope relationships between the genera Medi-
cago and Trigonella (Small 1988; Small et al. 1981a, b, c;
Small and Jomphe 1989). However, no strict delimitation
could be proposed.
Several cytological investigations have been conducted
on Trigonella (Singh and Roy 1970; Singh and Singh 1976;
Agarwal and Gupta 1983; Ahmad et al. 1999; Dundas et al.
2006; Aykut et al. 2009; Martin et al. 2008, 2010, 2011a,
b). Chromosome numbers of 45 taxa of Turkish Trigonella
belonging to 13 sections were determined as 2x = 14, 16,
30, and 46 by Martin et al. (2011a, b).
Seed coat characters have provided an approach to the
systematic relationships among the species of the Trigo-
nella (Small et al. 1989; Gupta 1991; Taia 2004a). Seeds
morphology of 37 taxa of Trigonella from various regions
in Turkey was examined with a scanning electron micro-
scope (SEM) and light microscope (LM). Based on seed
features such as shape, colour, and surface ornamentation
pattern, seven morphological types were recognized Ceter
et al. (2012).
Studies on the pollen grains of the family Fabaceae have
dealt mainly with the description of the pollen of certain
genera or sometimes tribes (e.g., Clarke and Kupicha 1976;
Ferguson and Skvarla 1979, 1981, 1982; Ferguson 1978,
1980, 1981; Ferguson and Strachan 1982; Diez and Fer-
guson 1994; Perveen and Qaiser 1998; Pinar et al. 2009a;
Ceter et al. 2013a, b). Pollen morphology of Melilotus,
Trifolium, Medicago, Factorovskya, and Trigonella of tribe
Trigonelleae was examined using LM and SEM by Small
et al. (1981a, b, c). Investigation on pollen–ovule patterns
(Small 1986) performed to resolve relationships within and
between these genera. A comparative study on the Egyp-
tian species of genus Trigonella showed the presence of ten
species belonging to five sections (Hassan 1992). Ming-Jou
and Tseng-Chieng (1999) studied the pollen morphology
and seed of Trigonella hamosa Linn. by SEM that repre-
sent as new record to the flora of Taiwan. Gazar (2003)
studied the pollen morphology of 28 species belonging to
the three genera (Melilotus, Trifolium and Trigonella) of
subfamily Papilionoideae in Egypt to establish the taxo-
nomic position of the species with in genera. Lashin (2006)
described the pollen grains of Egypt Trigonella foenum-
graecum L. and T. stellata Forssk. species by LM and
SEM. The pollen morphology of Tunisia T. foenum-grae-
cum, T. maritima Delile ex Poir., T. stellata and T. mons-
peliaca L. was studied by Haouala and Beji (2008). The
effect of maleic hydrazide (MH) has been studied on pollen
fertility of T. foenum-graecum to test its cytotoxicity (Jabee
et al. 2008). The pollen morphological characters of Egypt
T. foenum-graecum were investigated by the aid of LM and
SEM (El-Sayed et al. 2010). T. lasiocarpa Ranjbar and
Hajmoradi, T. binaloudensis Ranjbar and Karamian, T.
stipitata Ranjbar and Joharchi and T. torbatejamensis
Ranjbar, T. heratensis Rech. and T. adscendens Aphan. and
Gontsch from Iran are compared with their closest relatives
using morphology and pollen micromorphology (Ranjbar
et al. 2012b). The pollen morphology of T. yasujensis
Ranjbar, Hajmoradi, Karamian and T. elliptica Boiss. was
studied by Ranjbar et al. (2012a).
There are some other studies focusing on the Trigonella
species out of Turkey; (Sirjeav 1935), Meusel and Jager
(1962), Baum (1968), Small et al. (1981a, b), Classen et al.
(1982), Small (1988), Danin and Small (1989), Small and
Jomphe (1989), Al-Habori et al. (1998), Sheoran et al.
(1999), Ram and Verma (2000), Murakami et al. (2000),
Oncina et al. (2000), Sur et al. (2001), Sabir et al. (2002)
and Petropoulus et al. (Petropoulos 2002).The members of
Trigonella and particularly of the cultivated species T.
foenum–graecum (fenugreek) were known and used for
different purposes in ancient times, especially in Greece
and Egypt (Petropoulos 2002). In N. Africa, it has been
690 N. M. Pinar et al.
123
Author's personal copy
cultivated around the Saharan oases since very early times
(Duke 1986).
In this study, the pollen grains of 38 Trigonella taxa
showing morphological similarities were investigated with
LM and SEM, so that palynological comparisons could be
made and inter- and intra-generic relationships based on
pollen morphological data could be discussed. This study
attempts to clarify the systematics of this genus.
Materials and methods
Plant materials
Materials used for this study were collected from wild
populations. Collectors and localities are shown in the
specimens investigated list (Table 1). The specimens of the
plants are deposited in the Gazi (GAZI) and Harran (HRU)
Universities Herbaria. The order of the species was adapted
from Davis (1970) and Akan et al. (2005). All the speci-
mens are deposited in GAZI and HRU.
Palynological analysis
Pollen slides were prepared using the Wodehouse (1935)
technique. Pollen grains were stained with glycerin gel
with safranin, and slightly heating the slide and then a
coverslip was placed on it. The prepared slides were
studied under the light microscope. Their photographs were
taken with the Leica DM3000 digital photomicrograph
system (Germany). Measurements were based on 20 or
more pollen grains per specimen. For scanning electron
microscopy (SEM) studies, dried pollen grains were
transferred on aluminum stubs and coated with gold for
4 min in a sputter coater (Pinar et al. 2009b; Altuner et al.
2012; Ceter et al. 2013c). Morphological observations were
made in a Jeol JSM 6490LV Scanning electron microscope
at TPAO Research Center SEM Laboratory, Ankara.
Terminology was adopted from Faegri and Iversen
(1992), Punt et al. (1994, 2007), Hesse et al. (2009), Pınar
et al. (2009a), and shape classification follows that of Er-
dtman (1969) based on P/E ratio in Table 2.
Numerical analysis
The Simpson and Roe graphical test (van der Pluym and
Hideux 1997) was used for statistical calculations (Fig. 1).
Pollen characters of the taxa, coefficient of correlation
were determined, and they were grouped using the clus-
tering analysis method (UPGMA, dissimilarity, standard-
ized variable).
Table 1 The collections data of investigated Trigonella
Taxon Locality Voucher
T. cretica (L.) Boiss. C2 Burdur Akan & Ekici 3480
T. plicata Boiss & Bal. C4 Konya Akan, Aytac &
Ekici 3789
T. brachycarpa (Fisch.) Moris. C3 Antalya Akan & Ekici 3486
Trigonella lunata Boiss. C6 Adana Akan & Ekici 4687
T. corniculata L. C2 Mugla Akan & Ekici 2827
T. spinosa L. C2 Mugla Akan & Ekici 5655
T. coelesyriaca Boiss. C6
Gaziantep
Akan & Ekici 2610
T. spruneriana var. spruneriana
Boiss.
C3 Antalya Akan 3330
T. spruneriana var. sibthorpii
Fenzl.
C6 Antalya Akan & Ekici 3976
T. mesopotamica Desv. C8 Mardin Akan & Ekici 5578
T. cilicica C6 Adana Akan & Ekici 4682
T. filipes Boiss. C7
Sanlıurfa
Akan & Ekici 3200
T. velutina Boiss. A9 Kars Akan, Aytac &
Ekici 3743
T. strangulata Boiss. C8 Mardin Akan & Ekici 5577
T. smyrnea Boiss. C3 Antalya Akan & Ekici 4630
T. arenicola Hub.-Mor. C3 Antalya Akan & Ekici 3311
T. fischeriana Ser. B5 Kayseri Akan 3654
T. tenuis Fisch. A9 Kars Akan, Aytac &
Ekici 3742
T. astroites Fisch & Mey. B5 Aksaray Akan & Ekici 4750
T. halophila Boiss. C5 Mersin Akan & Ekici 4555
T. crassipes Boiss. B2 Denizli Akan & Ekici 3452
T. rigida Boiss.& Bal. C6 Adana Akan & Ekici 4737
T. pamphylica C3 Antalya Akan & Ekici 3331
T. carica Hub. Mor. C2 Mugla Akan & Ekici 3366
T. monantha C.A. Meyer subsp.
monantha
C2 Burdur Akan & Ekici 3466
T. orthoceras A9 Kars Akan & Ekici 3738
T. monspeliaca L. A7
Trabzon
Akan & Ekici 3690
T. isthmocarpa Boiss. & Bal. B5 Aksaray Akan & Ekici 4751
T. rhytidocarpa Boiss. & Bal. C5 Nigde Akan & Ekici 3590
T. spicata Sibth & Sm. A5 Samsun Akan & Ekici 4715
T. cephalotes C2 Mugla Akan & Ekici 2830
T. procumbens B3 Konya Akan & Ekici 4650
T. capitata Boiss. C2 Denizli Akan & Ekici 4767
T. coerulescens (Bieb.) Hal. C1 Aydın Akan, Aytac &
Ekici 3439
T. gladiata Stey Fischer B1 Izmir, Akan & Ekici 4622
T. foenum-graecum Boiss. C6
Gaziantep
Akan & Ekici 3227
T. macrorrhyncha Boiss. C5 Mersin Akan & Ekici 3293
T. raphanina Boiss. C5 Icel Akan & Ekici 2848
Comparative pollen morphology of annual Trigonella L. (Fabaceae) 691
123
Author's personal copy
Five palynological (consisting of nominal and qualita-
tive) characters were selected to distinguish the 38 taxa
(OTUs) of the genus Trigonella. Five morphological
characters state and their values or scales are given at
Table 2. A primary mixed data matrix was created for 38
taxa (OTUs) and five palynological characters (Table 2)
for the analysis. Since Gower’s formula (Jabee et al. 1971)
modified by Podani (1999) allows the inclusion of ordinal
variables and missing scores in the data matrix, it was used
to calculate the primary mixed data for dissimilarities.
Unweighted pair group method using arithmetic averages
(UPGMA) was selected because it is the most commonly
used method (Mohammadi and Prasanna 2003) and it has
advantages to other methods in: accurate reflection of the
similarity matrix as measured by the co-phenetic correla-
tion coefficient of Sokal and Rohlf (1962), symmetrical
hierarchical structure [the ‘‘structural value’’ concept of
McNeill (1979)], and congruence with classification
derived by traditional methods (Ward 1993). All compu-
tations were made by the MVSP 3.1 software.
Table 2 Five palynological characters to distinguish the 38 taxa of the genera Trigonella
Taxa/character states T1 T2 T3 T4 T5 T6 T7 T8 T9 T10 T11 T12 T13 T14 T15 T16 T17 T18 T19
Equatorial orn. 0 0 1 1 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0
Polar orn. 0 0 1 1 2 2 0 2 2 2 0 2 2 2 2 0 0 0 0
Aperture orn. 0 1 2 2 0 0 1 0 0 0 0 0 0 0 0 1 1 1 1
Pollen shape 0 0 1 2 2 2 2 3 2 3 4 2 3 0 2 2 2 2 2
Clg 4.2 3.1 1.6 2.7 4.3 2.1 2.3 2.8 2.4 2.6 4.2 3.1 1.8 3.1 2.3 5.2 2.4 4.2 4.2
Taxa/character states T20 T21 T22 T23 T24 T25 T26 T27 T28 T29 T30 T31 T32 T33 T34 T35 T36 T37 T38
Equatorial orn. 2 0 2 0 2 2 0 0 0 0 3 3 0 0 3 0 0 0 0
Polar orn. 3 0 3 0 3 3 0 0 0 0 1 1 0 0 3 0 0 0 0
Aperture orn. 1 1 1 1 1 1 1 1 1 1 2 2 1 1 3 0 0 0 0
Pollen shape 2 0 0 2 0 4 0 0 2 0 2 2 2 5 3 3 3 3 3
Clg 2.1 4.2 4.7 7.8 3.2 4.2 5.2 5.2 4.2 5.2 3.1 4.2 5.3 1.6 2.1 2.1 2.6 2.8 3.6
Fig. 1 Simpson and Roe test
for Trigonella. a Polar axes (P),
b equatorial axes (E)
692 N. M. Pinar et al.
123
Author's personal copy
Table 3 Qualitative pollen features of different taxa of Trigonella (value in lm)
Taxa P/E Pollen shape Aperture type Ornamentation
Equatorial Polar
Sect: Samaroideae
T. cretica (T1) 1.07 Prolate-spheroidal Tricolporate Microreticulate Psilate-perforate
Sect: Pectinatae
T. plicata (T2) 1.06 Prolate-spheroidal Tricolporate Microreticulate Psilate-perforate
Sect: Lunatae
T. brachycarpa (T3) 0.87 Suboblate Tricolporate Rugulate-scabrate Rugulate-scabrate
T. sirjaevii (T4) 1.28 Subprolate Tricolporate Rugulate-scabrate Rugulate-scabrate
Sect: Falcatulae
T. corniculata (T5) 1.24 Subprolate Tricolporate Microreticulate Microreticulate
T. spinosa (T6) 1.2 Subprolate Tricolporate Microreticulate Microreticulate
T. coelesyriaca (T7) 1.18 Subprolate Tricolporate Microreticulate Psilate-perforate
Sect: Cylindricae
T. spruneriana var. spruneriana (T8) 1.72 Prolate Tricolporate Microreticulate Microreticulate
T. spruneriana var. sibthorpii (T9) 1.34 Prolate Tricolporate Microreticulate Microreticulate
T. mesopotamica (T10) 1.58 Prolate Tricolporate Microreticulate Microreticulate
T. cilicica (T11) 0.96 Oblate- spheroidal Tricolporate Microreticulate Psilate-perforate
T. filipes (T12) 1.3 Subprolate Tricolporate Microreticulate Microreticulate
T. velutina (T13) 1.4 Prolate 60 %Tricolporate
20 %Triporate
20 %Abnormal pollen
Microreticulate Microreticulate
T. strangulata (T14) 1.07 Prolate-spheroidal 10 %Trisyncolporate
90 %Tricolporate
Microreticulate Microreticulate
T. smyrnea (T15) 1.25 Subprolate 10 %Tricolpate
%90 Tricolporate
Microreticulate Microreticulate
Sect: Bucerates
T. arenicola (T16) 1.14 Subprolate 20 %Tricolpate
20 %Pantocolporate
60 %Trizonocolporate
Microreticulate Psilate-perforate
T. fischeriana (T17) 1.13 Subprolate Tricolporate Microreticulate Psilate-perforate
T. tenuis (T18) 1.15 Subprolate %80 Tricolporate
%20 Abnormal pollen
Microreticulate Psilate-perforate
T. astroites (T19) 1.32 Subprolate Tricolporate Microreticulate Psilate-perforate
T. halophila (T20) 1.15 Subprolate Tricolporate Perforate Perforate
T. crassipes (T21) 1.11 Prolate-spheroidal Tricolporate Microreticulate Psilate-perforate
T. rigida (T22) 1.07 Prolate-spheroidal Tricolporate Perforate Perforate
T. pamphylica (T23) 1.25 Subprolate Tricolporate Microreticulate Psilate-perforate
T. carica (T24) 1.14 Prolate-spheroidal Tricolporate Perforate Perforate
T. monantha subsp. monantha (T25) 0.9 Oblate-spheroidal Tricolporate Perforate Perforate
T. orthoceras (T26) 1.02 Prolate-spheroidal %1 Tricolpate
%5 Tetracolporate
%94 Tricolporate
Microreticulate Psilate-perforate
Sect: Reflexae
T. monspeliaca (T27) 1.11 Prolate-spheroidal Tricolporate Microreticulate Psilate-perforate
Sect: Isthmocarpae
T. isthmocarpa (T28) 1.13 Subprolate %80 Tricolporate
%20 Tricolpodiporate
Microreticulate Psilate-perforate
Comparative pollen morphology of annual Trigonella L. (Fabaceae) 693
123
Author's personal copy
Table 3 continued
Taxa P/E Pollen shape Aperture type Ornamentation
Equatorial Polar
T. rhytidocarpa (T29) 1.22 Prolate-spheroidal %80 Tricolporate
%20 Tricolpodiporate
Microreticulate Psilate-perforate
Sect: Uncinatae
T. spicata (T30) 1.3 Subprolate Tricolporate Rugulate-scabrate Rugulate-scabrate
T. cephalotes (T31) 1.15 Subprolate Tricolporate Rugulate-scabrate Rugulate-scabrate
Sect: Capitatae
T. procumbens (T32) 1.2 Subprolate Tricolporate Microreticulate Psilate-perforate
T. capitata (T33) 1 Spheroidal Tricolporate Microreticulate Psilate-perforate
Sect: Biebersteinianae
T. coerulescens (T34) 1.45 Prolate Tricolporate Retipilate Perforate
Sect: Foenum-graecum
T. gladiata (T35) 1.39 Prolate Tricolporate Microreticulate Psilate-perforate
T. foenum-graecum (T36) 1.5 Prolate Tricolporate Microreticulate Psilate-perforate
T. macrorhyncha (T37) 1.67 Prolate Tricolporate Microreticulate Psilate-perforate
T. raphanina (T38) 1.39 Prolate Tricolporate Microreticulate Psilate-perforate
* Martin et al. (2008, 2011a, b)
Table 4 Quantitative pollen features of different taxa of Trigonella (value in lm)
Taxa Chromosome number (2n)/
ploidy level (x)*
Polar axis (P) Equatorial axis (E) Exine Intine Colpus Pore
Min Max Mean Min Max Mean Clt Clg Plt Plg
Sect: Samaroideae
T. cretica (T1) 16 (2x) 24.9 31.2 27.8 22.9 31.2 26 1.8 0.3 4.2 22.4 8.3 7.3
Sect: Pectinatae
T. plicata (T2) 14 (2x) 26 32.2 28.6 22.9 31.2 27.1 1 0.3 3.1 23.9 10.9 8.8
Sect: Lunatae
T. brachycarpa (T3) 16 (2x) 13.5 20.8 17.2 16.6 22.9 19.8 2.4 0.6 1.6 12.5 4.1 2.3
T. sirjaevii (T4) 16 (2x) 15.6 24.8 22.3 15.6 22.9 17.4 2.1 0.5 2.7 18.4 7.4 7.2
Sect: Falcatulae
T. corniculata (T5) 16 (2x) 15.2 29.1 23.1 16.6 21.8 18.6 1.1 0.3 4.3 17.4 5.4 6.7
T. spinosa (T6) 16 (2x) 13.5 18.7 15.6 10.4 16.6 13 0.8 0.3 2.1 12 4.2 2.6
T. coelesyriaca (T7) 16 (2x) 20.8 29.1 24.6 17.7 26 20.8 0.8 0.3 2.3 18.7 9.4 7.3
Sect: Cylindricae
T. spruneriana var.
spruneriana (T8)
16 (2x) 26 32.2 28.6 14.6 20.8 16.6 0.6 0.3 2.8 22.9 5.2 4.2
T. spruneriana var.
sibthorpii (T9)
16 (2x) 15.2 22.9 18.7 11.4 20.8 15.1 0.8 0.5 2.4 15.4 5.4 4.3
T. mesopotamica (T10) 16 (2x) 26 31.2 27.4 14.5 20.8 17.3 0.8 0.3 2.6 22.9 8.3 4
T. cilicica (T11) 16 (2x) 20.8 31.2 23.4 17.6 30.1 24.2 1.4 0.5 4.2 19.7 6.7 4.2
T. filipes (T12) 16 (2x) 25 33.3 29.8 19.8 25.2 22.9 1.3 0.3 3.1 26 6.2 6.2
T. velutina (T13) 16 (2x) 21.4 26 23.9 14.5 20.8 17.7 0.8 0.3 1.8 23.4 7.8 5.2
T. strangulata (T14) 16 (2x) 29.1 34.3 31.2 26 34.3 29.1 1.3 0.3 3.1 26 8.3 6.2
T. smyrnea (T15) 16 (2x) 20.8 29.1 26 16.6 26 20.8 1.1 0.3 2.3 23.9 2.6 1.8
Sect: Bucerates
T. arenicola (T16) 16 (4x) 30.2 41.6 34.8 22.9 35.4 30.4 1.8 0.6 5.2 31.2 6.2 11.4
T. fischeriana (T17) 14 (4x) 31.2 42.6 37.2 29.1 36.4 32.8 1.4 0.5 2.4 28.8 8.8 7.8
T. tenuis (T18) 16 (6x) 21.8 31.2 24.4 18.7 23.9 21.3 0.8 0.3 4.2 19.8 7.3 7.3
694 N. M. Pinar et al.
123
Author's personal copy
Results
Detailed pollen morphological features of the investigated
taxa are summarized in Tables 3, 4 and representative
pollen grains are illustrated in Figs. 2, 3, 4, 5, 6, 7, 8, 9, 10.
Size, symmetry and shape
The pollen grains of Trigonella are radially symmetrical
and isopolar. The pollen grains of the Trigonella are pro-
late-spheroidal, oblate-spheroidal, suboblate, spheroidal,
and prolate with the polar axes 13.5–49.9 lm and the
equatorial axes 10.4–43.7 lm (Fig. 1). Their dimensions
are smaller in T. spinosa and T. brachycarpa, and larger in
T. carica and T. rhytidocarpa. Their outlines are oblong in
equatorial view and circular in polar view: amb semicir-
cular. (Tables 3, 4; Figs. 2, 3, 4, 5, 6, 7, 8, 9, 10).
Apertures
The pollen grains of Trigonella are usually tricolporate or
rarely tricolpate, tripantocolporate, tetracolporate, and
colpodiporate. Some species have shown heteromorphic
characteristics. For example, 10 % tricolpate and 90 %
tricolporate in T. smyrnea, 20 % tricolpate, 20 % panto-
colporate and 60 % trizonocolporate in T. arenicola, 1 %
tricolpate, 5 % tetracolporate and 94 % tricolporate in T.
orthoceras, 20 % tricolpodiporate and 80 % tricolporate in
T. isthmocarpa. Colpus is short or long (12–39.5 lm) and
narrow or broad (1.6–7.8 lm) and por is circular or
lalongated. The highest values were observed in T. carica
and T. rhytidocarpa. T. spinosa and T. brachycarpa have
smallest values of colpus. Margins distinct are regular and
ends are acute. Colpus membrane is more or less granulate
(Tables 3, 4; Figs. 2, 3, 4, 5, 6, 7, 8, 9, 10).
Table 4 continued
Taxa Chromosome number (2n)/
ploidy level (x)*
Polar axis (P) Equatorial axis (E) Exine Intine Colpus Pore
Min Max Mean Min Max Mean Clt Clg Plt Plg
T. astroites (T19) 16 (4x) 29.1 34.3 31.7 20.8 28.1 23.9 1.3 0.3 4.2 26 8.3 6.2
T. halophila (T20) 16 (4x) 29.1 36.4 32.2 21.8 33.3 28.1 1.3 0.3 2.1 26 10.9 9.9
T. crassipes (T21) 16 (4x) 25 32.2 28.1 22.9 28.1 25.3 1.7 0.5 4.2 19.8 6.2 6.2
T. rigida (T22) 16 (4x) 30.2 36.4 32.2 25 36.4 30.2 2.1 0.6 4.7 25 11.4 9.4
T. pamphylica (T23) 16 (4x) 36.4 42.6 39.5 29.1 34.3 31.7 1.3 0.8 7.8 31.2 10.4 9.9
T. carica (T24) 16 (4x) 42.6 49.9 46.3 38.4 43.7 40.6 2.3 1 3.2 36.4 17.7 12.5
T. monantha subsp.
monantha(T25)
16,28, 30 (4x) 26 33.3 29.1 29.1 36.4 32.2 1.6 0.3 4.2 23.4 5.7 7.3
T. orthoceras (T26) 46 (4x) 23.9 31.2 27.1 22.9 31.2 26.5 1.1 0.3 5.2 23.9 7.3 6.2
Sect: Reflexae
T. monspeliaca (T27) 16 (2x) 21.8 31.2 26.5 20.8 27.1 23.9 1.4 0.6 5.2 18.7 6.2 5.2
Sect: Isthmocarpae
T. isthmocarpa (T28) 16 (2x) 29.1 36.4 32.3 25 32.2 28.7 1.6 0.5 4.2 29.1 4 4
T. rhytidocarpa (T29) 16 (2x) 39.5 47.8 43.2 29.1 39.5 35.4 3.1 0.6 5.2 39.5 10.4 10.4
Sect: Uncinatae
T. spicata (T30) 16 (2x) 22.9 31.2 26 17.7 23.9 20.8 1.3 0.3 3.1 21.8 9.4 6.2
T. cephalotes (T31) 16 (2x) 20.8 29.1 25 18.7 25 21.8 1.1 0.3 4.2 25 6.8 4.7
Sect: Capitatae
T. procumbens (T32) 18 (2x) 18.7 34.3 27.4 15.6 27.1 22.8 0.5 0.3 5.3 24.3 8.3 7.3
T. capitata (T33) 16 (2x) 16.6 22.9 19.8 16.6 21.8 19.8 1.3 0.6 1.6 15.6 6.2 5.1
Sect: Biebersteinianae
T. coerulescens (T34) 16 (2x) 29.1 40.6 33.3 18.7 26 22.9 2.1 0.3 2.1 26 7.3 7.3
Sect: Foenum-graecum
T. gladiata (T35) 16 (2x) 38.5 31.2 34.3 22.9 28.1 24.6 1.3 0.5 2.1 27.6 8.3 6.2
T. foenum-graecum
(T36)
16 (2x) 28.1 33.4 31.2 18.7 23.9 20.8 1.8 0.6 2.6 23.9 7.3 5.2
T. macrorhyncha (T37) 16 (2x) 43.7 47.8 52 28.1 33.3 31.2 2.8 0.4 2.8 35.9 13 7.8
T. raphanina (T38) 16 (2x) 39.5 46.8 42.6 27.1 34.3 30.7 1.8 0.3 3.6 31.7 10.4 8.3
* Martin et al. (2008, 2011a, b)
Clg colpus length, Clt colpus width, Plg porus length, Plt porus width
Comparative pollen morphology of annual Trigonella L. (Fabaceae) 695
123
Author's personal copy
Exine
The overall thickness of stratified exine ranges from 0.5- to
3.1-lm Ectexine is thicker than endexine. The intine
thickness ranges from 0.3 to 1.3 lm (Tables 3, 4). Various
ornamentation types were observed: microreticulate (sec-
tion Samaroideae, Pectinatae, Falcatulae, Cylindricae,
Bucerates, Reflexae, Isthmocarpae, Capitatae and
Fig. 2 Light microscope microphotograph of Trigonella. 1–4 T. cretica, 5–8 T. plicata, 9–12 T. brachycarpa, 13–16 T. sirjaevii, 17–20 T.
corniculata, 21–24 T. spinosa
696 N. M. Pinar et al.
123
Author's personal copy
Foenum-graecum), rugulate, scabrate (section Lunatae,
Uncinatae), perforate (T. halophila, T. rigida and T. carica
from section Bucerates) and retipilatae (section Bieber-
steinianae) at equator and psilate-perforate (section
Samaroideae, Pectinatae, Bucerates, Reflexae,
Isthmocarpae, Foenum-graecum, T. coelesyriaca, T. cili-
cica), rugulate-scabrate (section Lunatae, Uncinatae),
microreticulate (Falcatulae, Cylindricae and Foenum-
graecum) and retipilatae (section Biebersteinianae) at polar
region. The lumina in the microreticulate regular and
Fig. 3 Light microscope microphotograph of Trigonella. 1–4 T. coelesyriaca, 5–8 T. spruneriana var. spruneriana, 9–12 T. spruneriana var.
sibthropii, 13–16 T. mesopotamica, 17–20 T. cilicica, 21–24 T. filipes
Comparative pollen morphology of annual Trigonella L. (Fabaceae) 697
123
Author's personal copy
0.3–1.5 lm diameter over the equatorial surface and muri
usually deep, concave and 0.075–0.13 lm thickness. The
ornamentation near aperture varies between microreticulate
in section Samaroideae, Falcatulae, Cylindricae, and
Foenum-graecum and psilate in section Pectinatae,
Bucerates, Reflexae, Isthmocarpae, Capitatae,
Fig. 4 SEM microphotograph of Trigonella. 1, 2 T. cretica, 3, 4 T.
plicata, 5, 6 T. brachycarpa, 7, 8 T. sirjaevii, 9, 10 T. corniculata, 11,12 T. spinosa, 13, 14 T. coelesyriaca, 15, 16 T. spruneriana var.
spruneriana, 17, 18 T. spruneriana var. sibthropii, 19, 20 T.
mesopotamica, 21, 22 T. cilicica, 23, 24 T. filipes
698 N. M. Pinar et al.
123
Author's personal copy
Biebersteinianae and T. coelesyriaca). In Trigonella,
numerical analysis of the differential palynological char-
acters, which became evident during the course of our
investigations, led to the realization of a dendrogram. This
dendrogram shows the similarities or dissimilarities which
exist among the taxa being studied. A dendrogram of
cluster analysis of Turkish Trigonella taxa based on five
character states of 38 taxa (OTUs) has been constructed.
Cluster analysis divided taxa into three main groups,
namely, cluster A, B and C. Cluster A includes:
Fig. 5 Light microscope microphotograph of Trigonella. 1–4 T. velutina, 5–8 T. strangulata, 9–12 T. smyrnea, 13–16 T. arenicola, 17–20T. fischeriana, 21–24 T. tenuis
Comparative pollen morphology of annual Trigonella L. (Fabaceae) 699
123
Author's personal copy
T. coerulescens, T. monantha subsp. monantha, T. carica,
T. rigida, T. halophila; Cluster B includes: T. spicata, T.
uncinata, T. brachycarpa, T. sirjaevii; Cluster C1 includes:
T. smyrnea, T. strangulata, T. velutina, T. filipes, T. mes-
opotamica, T. spruneriana var. spruneriana, T. spruneriana
var. sibthropii, T. corniculata, T. spinosa. Cluster C2
includes:T. cilicica, T. capitata, T. gladiata, T. foenum-
graecum, T. macrorhyncha, T. raphanina. Cluster C3
includes two sub-groups, namely, C3a and C3b. C3a
includes: T. coelesyriaca, T. arenicola, T. fischeriana,
Fig. 6 Light microscope microphotograph of Trigonella. 1–4 T. astroites, 5–8 T. halophila, 9–12 T. crassipes, 13–16 T. rigida, 17–20 T.
pamphylica, 21–24 T. carica
700 N. M. Pinar et al.
123
Author's personal copy
T. tenuis, T. astroites, T. pamphylica, T. isthmocarpa, T.
procumbens. C3b includes: T. cretica, T. pilicata, T.
crassipes, T. orthoceras, T. monspeliaca, T. rhytidocarpa.
Results presented according to palynological data in this
study showed that almost each of taxa separated between
own section (Fig. 11).
Fig. 7 SEM microphotograph of Trigonella. 1, 2 T. velutina, 3, 4 T. strangulata, 5, 6 T. smyrnea, 7, 8 T. arenicola, 9, 10 T. fischeriana, 11, 12T. tenuis, 13, 14 T. astroites, 15, 16 T. halophila, 17, 18 T. crassipes, 19, 20 T. rigida, 21, 22 T. pamphylica, 23, 24 T. carica
Comparative pollen morphology of annual Trigonella L. (Fabaceae) 701
123
Author's personal copy
Fig. 8 Light microscope
microphotograph of Trigonella.
1–4 T. monantha subsp.
monantha, 5–8 T. orthoceras,
9–12 T. monspeliaca, 13–16 T.
isthmocarpa, 17–20 T.
rhytidocarpa, 21–24 T. spicata,
25–28 T. cephalotes
702 N. M. Pinar et al.
123
Author's personal copy
Fig. 9 Light microscope
microphotograph of Trigonella.
1–4 T. procumbens, 5–8 T.
capitata, 9–12 T. coerulescens,
13–16 T. gladiata, 17–20 T.
foenum-graecum, 21–24 T.
macrorhyncha, 25–28 T.
raphanina
Comparative pollen morphology of annual Trigonella L. (Fabaceae) 703
123
Author's personal copy
Discussion
The grains of the taxa of Trigonella can be ascribed to the
‘‘Astragalus affghanus’’ type by Perveen and Qaiser
(1998), ‘‘1B and 2’’ types by Taia (2004b), on the basis of
pollen shape class (P/E) and exine sculpturing of type 1 by
Lashin (2006) and pollen shape class prolate type and
sculpture microreticulate subtype by El-Sayed et al. (2010).
Pollen grains of the genus Trigonella are heterogenous
and exhibit considerable variation in size (P and E) and
shape (P/E). (Tables 3, 4; Fig. 1). Similarly, Ceter et al.
(2012) found that the seeds of Trigonella are very variable
in size, shape, and colour. According to the values of the
seed length and width, the seeds are classified into four
groups. However, on the basis of pollen shape class (P/E),
there are three distinct pollen types viz., Type 1 (1.33
subprolate, prolate) Trigonella sp., Type 2 (1.22 subpro-
late) Melilotus, Medicago and Trifolium sp., Type 3 (1.11
prolate, spheroidal) Ononis sp. are recognized by Lashin
(2006). The thickness of exine and intine are almost sim-
ilar. (Tables 3, 4).
In general, speciation within Trigonella including
perennial species has occurred on the diploid level
(2n = 2x = 16) whereas with annuals are diploid, tetra-
ploid, and hexaploid with 2n = 2x = 16, 2n = 4x = 32
and 2n = 6x = 48 chromosome numbers (Martin et al.
2008; Aykut et al. 2009; Ranjbar et al. 2011). The annual
taxa of Turkish Trigonella have 2n = 14, 16, 30, and 46
chromosomes numbers (Martin et al. 2011a, b). Some
meiotic irregularities observed in different populations of
T. spruneriana and T. yasujensis included chromosomes
stickiness and formation of micronuclei in tetrad cells and
cytomixis (Ranjbar et al. 2011, 2012a, b). T. spruneriana
var. spruneriana (26–32.2 9 14.6–20.8 lm, prolate) and
T. spruneriana var. sibthorpii (15.2–22.9 9 11.4–20.8,
subprolate) show variation in size (P and E) and shape
(P/E) in Turkey. In addition, Chatuverdi et al. (1990) and
Brochman (1992) reported that pollen grain size strongly
correlates with the level of polyploid. Similarly, maximum
the values of P and E in tetraploid and hexaploid species of
section Bucerates have been observed. (Tables 3, 4;
Figs. 1, 2, 3, 4, 5, 6, 7, 8, 9, 10)
Based on tectum structure, four distinct pollen types
are distinguished in Melilotus sp. (tectate perforate,
reticulate and distinct pore), Medicago and Trigonella sp.
(tectate imperforate, reticulate, verrucate, and pore
indistinct), Trifolium sp. (semi-tectate, reticulate, and
pore indistinct) and Ononis sp. (tectate perforate, rugulate
and pore distinct) by Lashin (2006). Taia (2004b) recog-
nized that Trigonella has reticulate, foveolate, granulate
exine. We found that the pollen of annual Trigonella
members is very variable in exine ornamentation (mic-
roreticulate, rugulate-scabrate, perforate, retipilatae at
Fig. 11 Dendrogram showing dissimilarity distance of the examined taxa
Fig. 10 SEM microphotograph of Trigonella. 1, 2 T. monantha
subsp. monantha, 3, 4 T. orthoceras, 5, 6 T. monspeliaca, 7, 8 T.
isthmocarpa, 9, 10 T. rhytidocarpa, 11, 12 T. spicata, 13, 14 T.
cephalotes, 15, 16 T. procumbens, 17, 18 T. capitata, 19, 20 T.
coerulescens, 21, 22 T. gladiata, 23, 24 T. foenum-graecum, 25, 26 T.
macrorhyncha, 27, 28 T. raphanina
b
Comparative pollen morphology of annual Trigonella L. (Fabaceae) 705
123
Author's personal copy
equator region and psilate-perforate, rugulate-scabrate,
microreticulate, and retipilatae at polar region) (Tables 3,
4; Figs. 1, 2, 3, 4, 5, 6, 7, 8, 9, 10).
The general aperture form is tricolporate, but T. smyrnea
(10 % tricolpate and 90 % tricolporate), T. arenicola
(20 % tricolpate, 20 % pantocolporate, and 60 % trizono-
colporate), T. orthoceras (1 % tricolpate, 5 % tetracolpo-
rate, and 94 % tricolporate), and T. isthmocarpa (20 %
tricolpodiporate and 80 % tricolporate) show considerable
aperture type variation (Tables 3, 4; Figs. 1, 2, 3, 4, 5, 6, 7,
8, 9, 10, 11). Variations in pollen size and aperture type
were attributed to heteromorphy in pollen grains by Nair
and Kaul (1965) and Inceoglu (1973)
Acknowledgments The authors thank Scientific and Technological
Research Council of Turkey (TUBITAK, TBAG Project No. 2099)
for financial support.
References
Agarwal K, Gupta PK (1983) Cytological studies in the genus
Trigonella L. Cytologia 48:771–779
Ahmad F, Acharya SN, Mir Z, Mir PS (1999) Localization and
activity of rRNA genes on fenugreek (Trigonella foenum-
graecum L.) chromosomes by fluorescent in situ hybridization
and silver staining. Theor Appl Genet 98:179–185
Akan H, Ekici M, Aytac Z (2005) The endemic species of Trigonella
and their conservation in Turkey, XVII. International Botanical
Congress, Vienna, (17–23 July), Abstracts, p 611
Al-Habori M, Al-Aghbari AM, Al-Mamary M (1998) Effects of
fenugreek seeds and its extracts on plasma lipid profile: a study
on rabbits. Phytother Res 12(8):572–575
Altuner EM, Ceter T, Alpas H (2012) High hydrostatic pressure
processing: a method having high success potential in pollen
protein extraction. High Pres Res 32(2):291–298
Aykut Y, Martin E, Unal F, Akan H (2009) Karyological study on six
Trigonella L. Species (Leguminosae) in Turkey. Caryologia
6:89–94
Balodi B, Rao RR (1991) The genus Trigonella L. (Fabaceae) in the
northwest Himalaya. J Econ Tax But 5:11–16
Baum BR (1968) A classification of the generic limits of Trigonella
and Medicago. Canad J Bot 46:741–749
Bhatti MA, Khan MTJ, Ahmad B, Jamshaid M, Ahmad W (1996)
Antibacterial activity of Trigonella foenum-graecum seeds.
Fritoletapia 67:372–374
Brochman C (1992) Pollen and seed morphology of Nordic Draba
(Brassicacea): phylogenetic and ecological implications. Nord J
Bot 12:657–673
Ceter T, Karaman S, Aytac Z, Baser B (2013) Pollen morphology of
Oxytropis DC. (Fabaceae) genus in Turkey. Bangladesh J Bot
42(1):167–174
Ceter T, Pınar NM, Akan H, Ekici M, Aytac Z (2012) Comparative
seed morphology of Trigonella L. species (Leguminosae) in
Turkey. Afr J Agric Res 7(3):509–522
Ceter T, Ekici M, Pınar NM, Ozbek F (2013a) Pollen morphology of
Astragalus L. section Hololeuce Bunge (Fabaceae) in Turkey.
Acta Botanica Gallica 160(1):43–52
Ceter T, Pınar NM, Inceer H, Hayırlıoglu Ayaz S, Yaprak AE (2013b)
The comparative pollen morphology of genera Matricaria L. and
Tripleurospermum Sch. Bip. (Asteraceae) in Turkey. Plant Syst
Evol 299(5):959–977
Chatuverdi M, Yunus D, Nair PK (1990) Cytopalynological studies of
Arachis L. (Leguminosae). Cultivated and wild species and their
hybrids. Grana 29:109–117
Chopra RN, Nayar SL, Chopra IC, Asolkar LV, Kakkar KK, Chakre
OJ, Varma BS (1956) Glossary of Indian Medicinal Plants (1st
ed.)—CSIRO, New Delhi, p 248
Clarke GCS, Kupicha FLS (1976) The relationships of the genus
Cicer L. (Leguminosae): the evidence from polen morphology.
Bot J Linn Soc 72:35–44
Classen D, Nozzolillo C, Small E (1982) A phenolic–taxometric study
of Medicago (Leguminosae). Can J Bot 60:2477–2495
Dangi RS, Lagu MD, Choudhary LB, Ranjekar PK, Gupta VS (2004)Assessment of genetic diversity in Trigonella foenum–graecum
and Trigonella caerulea using ISSR and RAPD markers. BMC
Plant Biol 4:13–23
Danin A, Small E (1989) Contributions to the flora of Israel and
Sinae. V. Trigonella sibthorpii Boiss., a new record from Israel.
Israel J Bot 38:121–124
Davis PH (1970) Flora of Turkey and The East Aegean Islands.
Edinburgh Univ. Press, Edinburgh, pp 452–482
Diez MI, Ferguson LK (1994) Th e pollen morphology of the tribes
Loteae and Coronilleae, Lotus L. and related genera (Papilio-
noideae: Leguminosae). Rev Palaeobot Palynol 81:233–255
Duke AJ (1986) Handbook of Legumes of World Economic
Importance. Plemus Press, New York/London
Dundas IS, Nair RM, Verlin DC (2006) First report of meiotic
chromosome number and karyotype analysis of an accession of
Trigonella balansae (Leguminosae). N. Z. J Agric Res 49:55–58
El-Sayed SM, El-Sayed M, Tantawy A, Ghareeb I, El-Sheekh A.E
(2010) Pollen morphology of some species of subfamily
Papilionoideae. Proceedıng of Fifth Scientific Environmental
Conference, Zagazig Uni., 71–86
Erdtman G (1969) Handbook of Palynology, Morphology, Taxonomy
and Ecology. Munksgaard, Copenhagen
Faegri K, Iversen J (1992) Textbook of pollen analysis, 4th edn.
Wiley, New York
Ferguson IK (1978) A note on the pollen morphology of the genus
Cranocarpus Bentham (Leguminosae). Bradea 2(39):269–272
Ferguson IK (1980) The pollen morphology of the genus Ceratonia
(Leguminosae: Caesalpinoideae). Kew Bull 35
Ferguson IK (1981) The Pollen morphology of Macroyloma (Legu-
minosae: Phaseoleae). Kew Bull 36:455–761
Ferguson IK, Skvarla JJ (1979) The pollen morphology of Crano-
carpus marhi Bentham (Leguminosae: Papilionoideae). Grana
18:15–20
Ferguson IK, Skvarla JJ (1981) The pollen morpology of the
subfamily Papilionoideae (Leguminosae). In: Raven PH (ed)
RM Polhill. Advances in Legume systematics, Royal Botanic
Gardens, Kew, pp 859–896
Ferguson IK, Skvarla JJ (1982) Pollen morphology in relation to
Pollinators in Papilionoideae (Leguminosae). Bot J Linn Soc
84:183–193
Ferguson IK, Strachan R (1982) Pollen morphology and taxonomy of
the tribe Indigofereae (Leguminosae: Papilionoideae). Pollen
Spores 24:171–210
Gazar M (2003) Pollen morphology of the three genera of subfamily
Papilionoideae in Egypt (Melilotus, Trifolium and Trigonella).
Acta Botanica Hungarica 45(3):279–296
Girardon P, Sauvaire Y, Baccou JC, Bessiere JM (1986) Identifi
cation de la 3-hydroxy-4,5-dimethyl-2 (SH)-furanone dans
L’arome des grains de Fenugrec (Trigonella foenum-graecum
L.) Lebensm. Wiss Technol 19:44–46
Gupta M (1991) Seed coat structure in some species of Trigonella.
Scanning Microsc 5(3):787–796
Haouala R, Beji M (2008) Pollen morphology of some Tunisian
species from genus of Trigonella L. J Agric Sci Technol 2:6–11
706 N. M. Pinar et al.
123
Author's personal copy
Hassan AE (1992) Comparative studies on the Egyptian species of the
genus Trigonella. M.Sc. Thesis, Suez Canal Univ., Ismailia
Egypt
Hesse M, Halbritter H, Zetter R, Weber M, Buchner R, Frosch-
Radivo A, Ulrich S (2009) Pollen terminology, an ıllustrated
handbook. Springer-Verlag/Wien, Austria
Heyn CC (1981) Trifolieae. In: Polhill RM, Raven PH (eds) Advances
in legume systematics part 1. R Bot Gard, Kew, pp 383–385
Hutchinson J (1964) The Genera of Flowering Plants, vol I.
Clarendon Press, Oxford
Inceoglu O (1973) Asyneuma canescens (W.K.) Griseb. & Schenk’in
polen morfolojisi ve heteremof polenler. Turk Biyoloji Derg
23:89–94
Jabee G, Ansari MYK, Danish Shahab JC (1971) A general
coefficient of similarity and some of its properties. Biometrics
27:857–871
Jabee F, Ansari MYK, Shahab D (2008) Studies on the effect of
maleic hydrazide on root tip cells and pollen fertility in
Trigonella foenum-graecum. Turk J Bot 32:337–344
Kawashty SA, Abdalla MF, Gamal El Din EM, Saleh NAM (1998)
The chemosystematics of Egyptian Trigonella species. Biochem
Syst Ecol 26:851–856
Lashin GM (2006) Comparative morphology of pollen grains of some
taxa of tribe Trifolieae (Fabaceae: Papilionoideae) from Egypt.
Int J Bot 2(3):210–211
Lock JM, Simpson K (1991) Legumes of west Asia. R. Bot. Gard,
Kew
Mabberley DJ (1997) The plant book, a portable dictionary of the
vascular plants, 2nd ed. Cambridge Univ. Press
Martin E, Akan H, Ekici M, Aytac Z (2008) Karyomorphological
studies on section Bucerates Boiss. of Trigonella L. (Legumi-
nosae) from Turkey. Caryologia 61:225–236
Martin E, Akan H, Ekici M, Aytac Z (2010) Karyology of ten Turkish
Trigonella L. (Leguminosae) species from section Cylindricae
Boiss. Turkish J Bot 34:485–494
Martin E, Akan H, Ekici M, Aytac Z (2011a) New chromosome
numbers in the genus Trigonella L. (Fabaceae) from Turkey. Afr
J Biotechnol 10:116–125
Martin E, Akan H, Ekici M, Aytac Z (2011b) Karyotype analyses of ten
sections of Trigonella (Fabaceae). Comp Cytogenet 5:105–121
McNeill J (1979) Structural value: a concept used in the construction
of taxonomic classifications. Taxon 28:481–504
Meusel H, Jager E (1962) Uber die Verbreitung einiger Papiliona-
cean–Gattun.–Kulturpflanze Beih 3: 249-262. Turkey/Distr. of
genera Leg.: Lotus, Medicago, Onobrychis, Ononis, Trifolium,
Trigonella/Germ
Ming-jou W, Tseng-Ching H (1999) Notes on the Flora of Taiwan
(34)-Trigonella hamosa Forssk. (Leguminosae). Taiwania
44(3):376–383
Mohammadi SA, Prasanna BM (2003) Analysis of genetic diversity in
crop plants. Salient statistical tools and considerations. Crop Sci
43:1235–1248
Murakami T, Kishi A, Matsuda H et al (2000) Medicinal foodstuffs.
17. Fenugreek seed. (3): structures of new furostanol–type
steroid saponins, trigoneosides 10a,10b,11b,12a,12b and 13a
from the seeds of Egyptian Trigonella foenum–graceum L.
Chem Pharm Bull 48(7):994–1000
Nair PK, Kaul KN (1965) Pollen grain in a gigantic of Rauwolfia
serpentine. Current Sci 34:256–257
Oncina R, Botia JM, Del Rio JA, Ortuno A (2000) Bioproduction of
diosgenin in callus cultures of Trigonella foenum–graecum L.
Departemento de Biologia Vegetal, facultad de Biologia, Uni-
versidad de Murcia, Campus de Espinardo, Murcia
Perveen A, Qaiser M (1998) Pollen Flora of Pakistan—VIII
Leguminosae (Subfamily: Papilionoideae). Turk J Bot 22:73–91
Petropoulos GA (2002) Fenugreek, The Genus Trigonella, Medicinal
and Aromatic Plants–Indutsrial Profiles. TJ International Ltd.,
Padstow
Pınar NM, Duran A, Ceter T, Tug GN (2009a) Pollen and seed
morphology of the Genus Hesperis L. (Brassicaceae) in Turkey.
Turk J Bot 33(2):83–96
Pınar NM, Ekici M, Aytac Z, Akan H, Ceter T, Alan S (2009b) Pollen
morphology of Astragalus L. sect. Onobrychoidei DC. (Faba-
ceae) in Turkey. Turk J Bot 33:291–303
Podani J (1999) Extending Gower’s coefficient of similarity to ordinal
characters. Taxon 48:331–340
Polhill RM, Raven PH (1981) Advances in legume systematic. R. Bot.
Gard, Kew
Punt W, Blackmore S, Nilsson S, Le Thomas A (1994) Glossary of
pollen and spore terminology. LPP Foundation, Utrecht
Punt W, Hoen PP, Blackmore S, Nilsson S, Le Thomas A (2007)
Glossary of Pollen and Spore Terminology. Rev Palaeobot
Palynol 143:1–81
Ram D, Verma JP (2000) Effect of level of phosporus and potash on
the performance of seed vield of ‘‘Pusa Early Bunching’’
fenugreek (Trigonella foenum–graceum). Indian J Agr SCI
70(12):866–868
Ranjbar M, Hajmoradi Z, Karamian R (2011) Cytogenetic study and
pollen viability of four populations of Trigonella spruneriana
Boiss. (Fabaceae) in Iran. J Cell Mol Res 3(1):19–24
Ranjbar M, Hajmoradi Z, Karamian R (2012a) A new species and
taxonomic studies Trigonella sect. Ellipticae (Fabaceae) in Iran.
Ann Bot Fennici 49:279–287
Ranjbar M, Karamian R, Hajmoradi Z, Joharchi MR (2012b) A
revision of Trigonella sect. Ellipticae (Fabaceae) in Iran. Nordic
J Bot 30:17–35
Sabir DM, Hussain FHS, Zewar JB (2002) Seperation and charac-
terisation of fenugreek seeds proteins. Deut lebensm-rundsch
98(1):14–16
Sheoran RS, Sharma HC, Niwas R et al (1999) Thermal efficiency of
fenugreek(Trigonella foenum–graceum) genotypes under differ-
ent sowing environments and phosporus levels. Indian J Agr SCI
69(12):830–832
Singh A, Roy RP (1970) Karyological studies in Trigonella,
Indigofera and Phaseolus. Nucleus 13:41–54
Singh A, Singh D (1976) Karyotype studies in Trigonella. Nucleus
19:13–16
Sirjaev G (1934) Generis Trigonella L. revisio critica. Pars II, 1. Spisy
Prir Fak. Masr. Univ., 192: 1–15. Turkey/Leg.: Trigonella Notes
on history &taxon
Sirjeav G (1935) Die Entwicklungsgeschichte der Gattung Trigonella
(Suppl. Ad ‘‘Generis Trigonella revisio critica’’). Bull Assoc
Russe Rech Sci Prague 2(9):135–162
Small E (1986) Pollen—ovule patterns in tribe Trifolieae (Legumi-
nosae). Plant Syst Evol 160:195–205
Small E (1987) A taxonomic study of the ‘‘medicagoid’’ Trigonella.
Can J Bot 65:1199–1211
Small E (1988) Pollen–ovule patterns in tribe Trifolieae (Legumino-
sae). Plant Syst Evol 160:195–205
Small E, Jomphe M (1989) A note on Trigonella (Melilotus) bicolor.
Can J Bot 67:1604–1606
Small E, Bassett IJ, Crompton CW (1981a) Pollen variation in tribe
Trigonelleae (Leguminosae) with special reference to Medicago.
Pollen Spores 23:295–320
Small E, Crompton CW, Brookes B (1981b) The taxonomic value of
floral characters in tribeTrigonelleae (Leguminosae), with spe-
cial reference to Medicago. Can J Bot 59:1578–1598
Small E, Lefkovitch LP, Brookes B (1981c) Remarkable asymmetries
in trifoliolate leaves with particular reference to Medicago. Can J
Bot 59:662–671
Comparative pollen morphology of annual Trigonella L. (Fabaceae) 707
123
Author's personal copy
Small E, Brookes B, Lassen P (1989) Circumscription of the genus
Medicago (Leguminosae) by seed characters. Can J Bot
68:613–629
Sokal RR, Rohlf FJ (1962) The comparison of dendrograms by
objective methods. Taxon 11:33–40
Sur P, Das M, Gomes A, Vedasiromoni JR, Sahu NP, Banarjee S,
Sharma RM, Ganguly DK (2001) Trigonella foenum graceum
(Fenugreek) Seed Extract as an Antineoplastic Agent. Phytother
Res 15(3):257–259
Taia WK (2004a) Palynological study within tribe Trifolieae (Legu-
minosae). Pakistan J Biol Sci 7(7):1303–1315
Taia WK (2004b) Tribe trifolieae: evidence from seed characters.
Pakistan J Biol Sci 7(7):1287–1302
Townsend CC, Guest E (1974) Flora of Iraq, vol 3. Ministry of
Agricult. and Agrar. Reform, Baghdad
Tutin TG, Heywood VH (1964) Flora Europaea, vol I, II. Cambridge
University Press, Cambridge
Van der Pluym A, Hideux M (1997) Applications d’une methodologie
quantitative a la palynologie d’Eryngium maritimum (Umbel-
liferae). Plant Syst Evol 127:55–85
Vasil’chenko IT (1953a) Bericht uber die Arten der Gattung,
Trigonella. Trudy Bot Inst Akad Nauk SSSR I, 10
Vasil’chenko IT (1953b) Obzor vidov roda Trigonella L. Trudy Bot
Inst Akad Naul SSSR ser. 1 Fl. Sist Vyss Rast 10:124–269
Ward JM (1993) Systematics of New Zealand Inuleae (Compositae–
Asteraceae)-2, A numerical phenetic study of Raoulia in relation
to allied genera. New Zeal J Bot 31:29–42
Wodehouse PP (1935) Pollen Grains. McGraw-Hill, New York
708 N. M. Pinar et al.
123
Author's personal copy