Plant Syst Evol DOI 10.1007/s00606-013-0919-2
ORIGINAL ARTICLE
Pollen morphology and evolution in the genus Orobanche L. s.l. and its allied genera (Orobancheae/Orobanchaceae) in Turkey Golshan Zare • Ali A. Do¨nmez • Emel O. Do¨nmez
Received: 26 July 2013 / Accepted: 17 September 2013 Ó Springer-Verlag Wien 2013
Abstract The pollen morphology of 120 samples of the tribe Orobancheae, representing four genera and 40 species of Cistanche, Diphelypaea, Orobanche and Phelipanche native to Turkey, has been studied by light and scanning electron microscopy. Pollen of the tribe Orobancheae is typically isopolar, radially asymmetrical, oblate spheroidal or prolate and belongs to one of the following basic pollen types: inaperturate, tricolpate and syncolpate. Also, pollen heteromorphism is widespread among pest species of Orobanche and Phelipanche. Pollen characters display considerable variation among genera and species, but some characters are significant at the levels of genera and sections. The results of the SEM study show that there are various exine ornamentation types in these genera. The obtained results support the division of the traditionally circumscribed genus Orobanche sensu lato into two genera, Phelipanche and Orobanche. The pollen morphological features are discussed on the basis of molecular phylogeny of the taxa. Keywords Orobanchaceae Pollen characters Pollen heteromorphism Pollen evolution
Introduction The family Orobanchaceae Vent. represents the largest monophyletic group comprising parasitic plants. Traditionally considered to be closely related to Scrophulariaceae Juss., Orobanchaceae, as redefined by Young et al. G. Zare (&) A. A. Do¨nmez E. O. Do¨nmez Department of Biology, Faculty of Science, Hacettepe University, 06800 Beytepe, Ankara, Turkey e-mail:
[email protected]
(1999), is a morphologically diverse family comprising mostly hemiparasitic species, which were formerly placed in Scrophulariaceae subfamily Rhinanthoideae, and exclusively holoparasitic species of Orobanchaceae in its traditional circumscription, i.e. Orobanchaceae s.str. (sensu Beck Von Mannagetta 1890, 1930). Orobanche is one of the largest genera of this family and is placed in tribe Orobancheae. Until recently, Orobanche L. sensu lato has been treated under a single genus. Under this concept, it comprises ca. 170 species (Uhlich et al. 1995), 30–35 % of which are present in Turkey (Gilli 1982; Davis et al. 1988; Zare 2012). Following the treatment of Beck Von Mannagetta (1930), most authors divide Orobanche into four sections: Orobanche, Trionychon Wallr., Gymnocaulis Nutt., and Myzorrhiza (Phil.) Beck. However, based on morphological and karyological differences and supported by recent molecular phylogenetic analyses, these taxa have been treated as separate genera; i.e. Orobanche L., Phelipanche Pomel, Aphyllon Mitchell (= Thalesia Britton), and Myzorrhiza Philippi (Sojak 1972; Holub 1977, 1990; Teryokhin et al. 1993; Bennett and Mathews 2006, Schneeweiss et al. 2004a, b; Carlo´n et al. 2005; WeissSchneeweiss et al. 2006; Park et al. 2008). As a holoparasitic plant, Orobanche has strongly reduced vegetative parts, thus possessing only a few morphological features suitable for taxonomical studies. Therefore, morphological characters pertaining to reproductive organs, such as floral, pollen and seed characters, provide most important diagnostic characters in this group. Due to the difficulty of reliable assessment of morphological characters on dried plant material, taxonomy of the genus is difficult and more problematic than in other taxa. Nevertheless, there were several taxonomically oriented studies including some on pollen of this
123
123
Ia
Ib
O. owerini
O. picridis
Sph
Ia
Ia
O. lutea
O. minor
Sph
Sph
Sph
Sph
Sph
O-sph
Sph
O-sph
Ib
O. gracilis
Sph
–
Ia
O. fuliginosa
O-sph
O. longibracteata
Ia
O. elatior
O-sph
Ia
IIIa
O. cumana
Sph
O. kurdica
Ib
O. crenata
Sph
Ia
IIIa
O. cernua
O-sph
Ia
Ia
O. caryophyllacea
O-sph
O. hadroantha
IIIa
O. camptolepis
Sph
O-sph
O. hederae
IIIa
O. anatolica
Sph
Ib
Ia
D. tourneforti
Orobanche alba
Sph
Ib
Diphelypaea coccinea
P-sph
II
Cistanche salsa
Shape
Type
Species name
Inaperturate
Inaperturate
Inaperturate
Inaperturate
Inaperturate
Inaperturate
Inaperturate
Inaperturate
Inaperturate
Inaperturate
Inaperturate
Syncolpate
Tricolpate
Inaperturat
Inaperturat
Syncolpate
Inaperturate Tricolpate
Inaperturate
Syncolpate
Inaperturate Tricolpate
Syncolpate
Inaperturate Tricolpate
Inaperturate
Inaperturate
Inaperturate
tetracolpate
Tricolpate
Aperture type
Scabrateverrucate
Scabrate
Scabrate
Scabrate
–
Scabrate
Scabrate
Scabrate
Scabrate verrucate
Scabrate
1.00
1.00
1.00
1.00
0.99
1.00
1.00
0.99
1.00
1.00
0.99
0.94
Scabrateverrucat
Scabrate
1.00
1.00
0.98
0.96
1.00
0.99
1
1
1.08
P/E
Scabrateverrucat
Scabrateverrucat
Scabrate
Scabrate verrucate
verrucate
Scabrate -
perforate
Scabrate-
Verrucate
Verrucate
Microreticulate
Ornamentation
23.02 ± 1.34
25.05 ± 2.09
24.29 ± 2.75
24.10 ± 2.09
24.54 ± 1.32
27.03 ± 4.75
23.37 ± 2.48
22.09 ± 2.57
25.33 ± 1.63
24.24 ± 1.4
26.53 ± 3.58
18.19 ± 1.33
23.75 ± 1.82
22.45 ± 1.77
28.28 ± 2.43
21.74 ± 1.97
26.54 ± 2.97
25.51 ± 2.34
25.35 ± 1.29
26.23 ± 2.08
29.1 ± 1.94
Polar axis (lm)
23.02 ± 1.34
25.05 ± 2.09
24.29 ± 2.75
24.10 ± 2.09
24.60 ± 1.95
27.03 ± 4.75
23.37 ± 2.48
22.12 ± 2.77
25.33 ± 1.63
24.21 ± 1.71
26.59 ± 3.59
19.20 ± 1.19
23.62 ± 2.72
22.45 ± 1.89
28.70 ± 3.43
22.43 ± 1.29
26.44 ± 3.03
25.64 ± 3.01
25.35 ± 1.29
26.23 ± 2.08
26.85 ± 1.69
Equatorial axis (lm)
1.17 ± 0.31
1.23 ± 0.21
1.10 ± 0.08
1.10 ± 0.17
1.20 ± 0.25
1.17 ± 0.10
0.93 ± 0.08
1.00 ± .13
1.22 ± 0.31
1.20 ± 0.30
0.95 ± 0.1
0.85 ± 0.15
1.02 ± 0.06
1.13 ± 0.12
1.07 ± 0.14
0.97 ± 0.07
1.03 ± 0.02
1.18 ± 0.15
1.09 ± 0.16
1.03 ± 0.18
0.95 ± 0.08
Exine thickness, (lm)
0.95 ± 0.15
1.02 ± 0.04
0.95 ± 0.05
0.99 ± 0.01
0.96 ± 0.08
1.01 ± 0.06
0.93 ± 0.04
0.94 ± 0.13
0.99 ± 0.01
1.00 ± 0
0.95 ± 0.04
0.83 ± 014
0.95 ± 0.04
0.97 ± 0.04
0.94 ± 0.08
0.88 ± 0.08
0.98 ± 0.20
0.99 ± 0.05
0.9 ± 0.1
0.93 ± 0.09
1.0 ± 0.1
Intine thickness (lm)
–
–
–
–
–
–
–
–
–
–
–
10.20 ± 2.62
–
4.75 ± 0.42
–
4.00 ± 0.28
5.95 ± 1.20
–
–
–
6.44 ± 2.24
Apocolpium diameter
Table 1 Collection data of Orobancheae specimens examined here from pollen morphological point of view: average ± standard deviation
–
–
–
–
–
–
–
–
–
–
–
15.80 ± 1. 22
–
18.58 ± 2.16
–
16.20 ± 2.05
18.71 ± 1.64
–
–
–
19.1 ± 2.64
Length of colpi (lm)
–
–
–
–
–
–
–
–
–
–
–
3.40 ± 0.90
–
3.11 ± 0.60
–
3.53 ± 0.65
3.11 ± 0.74
–
–
–
4.66 ± 0.7
Width of colpi (lm)
38
38
38
38
–
–
38
–
38–76
–
38
38
38
38
38
–
38
38
38
38
40
Polyploidy 2n
G. Zare et al.
Sph
Sph
Sph
O-sph
IIIb
II
II
II
II
Phelipanche aegyptiaca
P. arenaria
P. bungeana
P. caesia
P. cilicica
Sph
IIIb
P. schultzii
P-sph
Psph
O-sph
O-sph
Syncolpate
Inaperturate Tricolpate
Syncolpate
Inaperturate Tricolpate
Tricolpate
Tricolpate
Tricolpate
Tricolpate
Tricolpate
Tricolpate
Tricolpate
Tricolpate
Tricolpate
Tricolpate
Tricolpate
Tricolpate
Sincolpate
Inaperturate Tricolpate
Inaperturate
Inaperturate
Inaperturate
Inaperturate
Aperture type
Microreticulate
Microreticulate
Microreticulate
Microreticulate
Microreticulate
Microreticulate
Microreticulate
Microreticulate
Microreticulate
Microreticulate
Microreticulate
Microreticulate
Microreticulate
Microreticulate
Microreticulate
Scabrate
Scabrate
Scabrate
–
Ornamentation
1.02
1.03
0.96
0.96
0.96
0.92
1.01
0.97
1.00
0.92
1.00
0.94
1.00
1.00
1.04
1.00
1.00
1.00
1.00
P/E
Sph spheroidal, O-sph oblate spheroidal, P-sph prolate spheroidal
Chromosome number is based on Schneeweiss et al. (2004b) research results
IIIb
P. ramosa
O-sph
II
II
II
II
P. nana
P. nowackiana
P. oxyloba
O-sph
O-sph
II
P. mutelii
P. purpurea
P-sph
II
P. lavandulacea
Sph
II
II
P. coelestis
P. heldereichii
Sph
P-sph
Sph
Ia
Ia
O. reticulata
Sph
Sph
Shape
O. stocksii
–
Ia
O. palaestina
O. pubescens
Type
Species name
Table 1 continued
23.96 ± 2.46
23.53 ± 3.17
21.87 ± 1.42
22.13 ± 1.47
24.30 ± 1.56
20.03 ± 1.48
21.50 ± 2.07
22.44 ± 1.45
23.87 ± 1.88
22.29 ± 1.26
25.30 ± 1.67
22.65 ± 1.39
22.60 ± 1.64
27.27 ± 1.78
25.15 ± 1.74
27.10 ± 1.95
31.31 ± 2.65
22.96 ± 2.34
24.48 ± 2.34
Polar axis (lm)
23.44 ± 2.12
22.72 ± 1.80
22.67 ± 1.50
22.85 ± 2.11
25.20 ± 1.42
21.71 ± 2.91
21.21 ± 1.16
22.94 ± 1.68
23.67 ± 1.95
24.04 ± 1.80
25.30 ± 1.67
23.92 ± 1.89
22.60 ± 1.64
27.27 ± 1.78
24.08 ± 1.64
27.10 ± 1.95
31.31 ± 2.65
22.96 ± 2.34
24.48 ± 2.34
Equatorial axis (lm)
1.04 ± 0.06
0.98 ± 0.09
1.00 ± 0.01
1.08 ± 0.22
1.10 ± 0.21
1.07 ± 0.04
0.97 ± 0.06
1.00 ± 0.15
1.00 ± 0.08
1.02 ± 0.03
1.00 ± 0.30
1.00 ± 0
1.05 ± 0.15
0.89 ± 0.16
1.06 ± 0.07
1.00 ± 0.0
1.00 ± 0.28
1.02 ± 0.13
0.90 ± 0.20
Exine thickness, (lm)
0.98 ± 0.02
0.90 ± 0.12
1.00 ± 0.10
0.98 ± 0.06
1.00 ± 0
0.96 ± 0.01
0.97 ± 0.08
0.99 ± 0.01
0.96 ± 0.08
0.98 ± 0.02
1.01 ± 0.04
1.00 ± 0
0.93 ± 0.15
0.80 ± 0.20
0.98 ± 0.01
0.98 ± 0.06
0.94 ± 0.08
0.96 ± 0.07
0.85 ± 0.23
Intine thickness (lm)
9.74 ± 0.36
10.03 ± 0.72
10.49 ± 1.25
5.96 ± 0.79
5.10 ± 0.73
6.45 ± 0.75
7.84 ± 2.06
6.35 ± 1.62
5.87 ± 2.01
5.8 ± 2.40
7.55 ± 4.59
4.30 ± 1.15
9.48 ± 2.54
5.52 ± 1.20
9.75 ± 1.71
–
–
–
–
Apocolpium diameter
19.42 ± 2.01
18.06 ± 1.62
16.86 ± 1.20
17.40 ± 0.43
18.80 ± 2.04
18.14 ± 0.53
18.54 ± 1.13
18.4 ± 0.56
17.28 ± 1.58
17.35 ± 0.91
17.3 ± 0.28
17.50 ± 1.43
17.70 ± 1.11
17.40 ± 0.47
20.34 ± 1.04
–
–
–
–
Length of colpi (lm)
5.15 ± 0.07
4.23 ± 0.84
6.25 ± 0.73
3.46 ± 0.08
7.30 ± 1.25
5.26 ± 1.56
5.88 ± 0.84
4.60 ± 0.14
6.00 ± 1.56
4.4 ± 0.42
5.8 6 ± 2.35
4.20 ± 0.78
6.32 ± 1.19
5.08 ± 1.20
5.65 ± 1.5
–
–
–
–
Width of colpi (lm)
–
24
24
–
24
24
24
24
–
24
–
–
24
24
24
38
–
–
Polyploidy 2n
Pollen morphology and evolution in the genus Orobanche L. s.1. and its allied genera
123
G. Zare et al.
group (Abu Sbaih et al. 1994; Minkin and Eshbaugh 1989; Lu et al. 2007). These studies found that pollen morphology is an important character for elucidating taxonomic and evolutionary relationships in tribes Rhinantheae and Orobancheae. The taxonomic implications of pollen characters distinguishing between Orobanchaceae s.str. and Rhinanthoid taxa were discussed by Minkin and Eshbaugh (1989), who recognized the continuity between Orobanchaceae s.str. and Rhinanthoid taxa. Also, Abu Sbaih et al. (1994) indicated that pollen characters are significant in identification of some group of taxa in the genus Orobanche s.l.. In the course of a revision for Orobanche in Turkey aiming at finding reliable characters for classification of these taxa, their pollen morphology has been studied. Turkey is home to about 40 species of the tribe Orobancheae: 22 species of Orobanche L., 15 species of Phelipanche Pomel, two species of Diphelypaea Nicolson and one species of Cistanche Hoffmg. (Gilli 1971, 1982; Zare et al. 2010; Zare 2012; Zare and Do¨nmez 2013). Most of these species have never been investigated palynologically. Therefore, in this study, we re-considered their potential value for the classification of Orobanche s.l. and related genera and compared the results with molecular phylogenetic analysis and traditional taxonomy.
Materials and methods Pollen samples of 120 samples from four genera of Orobancheae (taxonomy follows Zare 2012) used for this study was either collected in the field during the summer of 2008–2012 or taken from herbarium specimens. Voucher specimens of the plants are deposited at the Herbarium of Hacettepe University (HUB), Turkey. A list of the voucher specimens is given in Appendix 1. Pollen terminology follows Hesse et al. (2009). The pollen grains of some taxa were acetolyzed according to the method of Erdthman (1969). However, this method is unsuitable for Orobancheae pollen observation because it causes the exine of pollen grains to rupture. Therefore, we used Wodehouse (1935) method and embedded the pollen in glycerine jelly stained with safranin. Pollen was examined with a Leica DM 4000B light microscope and photographed with a DFC 320 camera. Polar axis (P), equatorial axis (E), colpus length, colpus width, exine and intine thickness were measured for 30 pollen grains using an eyepiece micrometer under 100 magnifications. The measurements were then averaged to get approximate sizes of the pollen for each taxon. The
123
Fig. 1 Pollen grains of the tribe Orobancheae (LM); a–c Cistanche c ¨ ztekin1491): a tricolpate grain in polar view; b tricolpate salsa (M. O grain in equatorial view; c tetracolpate grain in equatorial view. d, e Diphelypaea coccinea (AAD10686): inaperturate grain. f–i inaperturate grain: f O. minor (AAD4366); g O. gracilis (AAD 16798); h O. lutea (GZ25), inaperturate grain with trace of colpi; i O. pubescens (GZ360), inaperturate grain with trace of colpi. j–n polymorphic pollen grains in the section Inflatae: j O. cernua (GZ131), inaperturate grain; k O. cumana (GZ492), tricolpate grain in equatorial view; l tricolpate grain in polar view; m tricolpate grain in equatorial view; n O. camptolepis (GZ553), dicolpate grain in equatorial view. o, p O. anatolica (GZ419): o tricolpate grain in equatorial and polar view; p tricolpate grain in equatorial view. q, r O. oxyloba (GZ643): q tricolpate grain in equatorial and polar view; r tricolpate grain in equatorial view. s P. aegyptiaca (GZ347): tricolpate grain in equatorial and polar view. t P. ramosa (GZ333): dicolpate grain in equatorial view. Scale bars 20 lm
ranges of P length for species and for populations were illustrated by Box-plots generated in R (version 2.15.2). For SEM study, the grains were mounted on stubs and sputter-coated with gold–palladium. SEM examination was carried out using a Zeiss EVO 50 EP microscope.
Results The pollen morphology showed remarkable variation among genera. Therefore, we prefer to give a general description for the genera with reference to particular species, where necessary. The major pollen features of all investigated taxa are summarized in Table 1. Representative pollen grains and selected LM and SEM micrographs are illustrated in Figs. 1, 2, 3, 4, 5 and 6. Pollen are shed as monads, and their size varies from small to medium (P = 18.19–28.28 lm). Orobanche cumana Wallr. possess the smallest pollen grains, while O. reticulata Wallr. has the largest pollen grains (Table 1). The shape of pollen grains in equatorial view ranges from (P/E = 0.92–1.04) oblate spheroidal to spheroidal or prolate spheroidal (Table 1). The outline in polar view is circular or triangular (Fig. 1). The examined taxa show three main pollen types: inaperturate; tricolpate; di- or trisyncolpate (Fig. 1). Some species show intraspecific variation with respect to apertures. Simple colpi are distributed symmetrically. Colpi lengths are 15.80–20.34 lm. Colpus length is correlated with length of the P. Colpi ends were subobtuse (Fig. 1). Exine ornamentation shows variation between taxa and varies between scabrate, verrucate to microreticulate (Figs. 2, 3, 4, 5 and 6). Sexine and nexine could not be distinguished. Comparisons of polar and equatorial axes lengths between all species from the four genera and between the
Pollen morphology and evolution in the genus Orobanche L. s.1. and its allied genera
123
G. Zare et al.
Fig. 2 Pollen grains of the genera Cistanche and Phelipanche (SEM) (dehydrated pollen); tricolpate grains, in equatorial view and their exine ornamentation respectively: a, b Cistanche salsa (BM3075); c, d P. ramosa (GZ560); e, f P. nana (GZ381); g, h P. mutelii (GZ625b); i, j P. oxyloba (GZ623); k, l P. lavandulacea (GZ514); m,
123
n P. heldereichii (485): o, p P. coelestis (GZ654); q, r P. cilicica (IE2352); s, t P. bungeana (GZ467); u, v P purpurea (GZ495); w, x P. nowackiana (BM10929). Scale bars 3 lm (a, c, e, g, i, k, m, o, q, s, u, w) and 1 lm (b, d, f, h, j, l, n, p, r, t, v, x)
Pollen morphology and evolution in the genus Orobanche L. s.1. and its allied genera
Fig. 3 Pollen grain of some members of Phelipanche (dehydrated pollen): a P. schultzii (GZ582) tricolpate grain in equatorial view; b– f P. aegyptiaca (GZ625a): b tricolpate grain in oblique polar view; c,
d inaperturate pollen irregularly infolded; e, f exine ornamentation. Scale bars 1 lm
different populations of Orobanche and Phelipanche indicate size variation between the examined taxa. As polar and equatorial axes lengths showed the same patterns, we only show results for the P lengths (Figs. 7, 8 and 9).
Phelipanche
Pollen grains medium in size (P length 29.10 lm); radially symmetric; prolate spheroidal (P/E = 1.08); tricolpate or rarely tetracolpate with microreticulate exine ornamentation (Figs. 1, 2). Exine thickness 0.95 ± 0.08 lm and intine thickness 1.0 lm (Table 1).
Pollen grains small to medium; radially symmetrical, typically tricolpate, oblate spheroidal to prolate spheroidal (P/ E = 0.92–1.04) with a microreticulate or verrucate exine ornamentation (Fig. 3). The smallest pollen was found in P. nana (No¨e) Sojak (P 9 E) 20.03 9 21.71 lm and the largest one was found in P. arenaria (Borkh.) Pomel (P 9 E) 27.27 9 27.27 lm. Exine thickness 0.89–1.08 lm and intine thickness is 0.80–1.01 lm (Table 1). P. aegyptiaca Pomel, P. ramosa (L.) Pomel and P. schultzii Pomel show a high diversity in apertures, because tricolpate, syncolpate and inaperturate pollen grains are present (Figs. 1, 2 and 3).
Diphelypaea
Orobanche
Pollen grains small to medium in size (P 25.35–26.23 lm); asymmetric; spheroidal (P/E = 1); inaperturate with verrucate exine ornamentation (Figs. 1, 4). Exine thickness 1.09–1.18 lm and intine thickness 0.90 lm (Table 1). No significant differences have been observed between the two species, Diphelypaea coccinea (M.Bieb.) Nicolson and D. tournefortii (Desf.) Nicolson.
Pollen grains small to medium; generally asymmetric; inaperturate or syncolpate with scabrate or verrucate exine ornamentation (Figs. 1, 4, 5 and 6); the sculpture is mostly scabrate, but is verrucate in O. gracilis Smith, O. picridis FW.Schultz and O. crenata Forsskal. Some of the studied taxa, such as O. anatolica Boiss. et Reut. and O. camptolepis Boiss. & Reut., showed both the exine
Pollen morphology of the genera Cistanche
123
G. Zare et al.
Fig. 4 Pollen grain in the genus Diphelypaea and Orobanche anatolica (SEM, dehydrated pollen): a–c Diphelypaea coccinea, inaperturate pollen and exine ornamentation. d–i O. anatolica: d,
e dicolpate pollen in equatorial view; f inaperturate pollen; g tricolpate grain in polar view; h, i (GZ419–GZ575) exine ornamentation. Scale bars 1 lm (a, b, c, h, i) and 2 lm (d, e, f, g)
ornamentation types in different populations (Figs. 5, 6). Pollen grain shape is oblate spheroidal to spheroidal (P/ E = 0.94–1.00; Table 1). Pollen size was small and ranged from (P 9 E) 18.19 9 19.20 lm in O. cumana to 28.28 9 28.70 lm in O. reticulata. Exine thickness is 0.85–1.23 lm, Intine thickness is 0.85–1.02 lm (Table 1).
pollen characters with both traditional and phylogenetic classifications of the taxa is discussed below. Pollen of the studied taxa can be grouped into three basic types based on pollen aperture type. These three groups can be further subdivided into two minor groups each based on the basic exine ornamentation:
Discussion
Type I. Inaperturate and asymmetrical pollen (Orobanche and Diphelypaea). I a. exine ornamentation scabrate.
Pollen morphology Pollen morphology provides some valuable information for the taxonomy of Orobanchaceae (Abu Sbaih et al. 1994; Minkin and Eshbaugh 1989; Shavvon and Mehrvarz 2010). The pollen characters, which were most useful in characterizing the investigated genera of Orobancheae, include aperture number and exine ornamentation. Based on the species studied here, the congruence of some important
123
Fig. 5 Pollen grains of the genus Orobanche scanning electron c micrographs, inaperturate pollen showing the depression due to dehydration (irregularly infolded) (dehydrated pollen): a O. lutea (GZ470). b–e O. caryophyllacea (FT1007): b, c without trace of colpi; d, e with trace of colpi (arrows). f O. kurdica (FT1008) with trace of colpi. g–h O. picridis (GZ363). i O. fuliginosa (AAD14886). j O. hadroantha (GZ697). k O. crenata (GZ626). l O. hederae (GZ463). m O. pubescens (GZ352). n O. owerini (GZ477). o O. alba (GZ593). Scale bars 3 lm (a, b, d, f, g, i, l, m, o) and 1 lm (c, e, h, k, n)
Pollen morphology and evolution in the genus Orobanche L. s.1. and its allied genera
123
G. Zare et al.
Fig. 6 Polymorphic pollen grains in the genus Orobanche, section Inflatae (SEM) scanning electron micrographs (dehydrated pollen): a O. cernua (GZ574) inaperturate pollen. b O. camptolepis (GZ690) tricolpate pollen in equatorial view. c O. cernua (Moazzeni) syndicolpate pollen in equatorial view. d–f O. camptolepis
(GZ553): d inaperturate pollen; e, f exine ornamentation in O. cumana (GZ492). g O. cernua, tricolpate pollen in polar view. h O. cumana (GZ492). i O. cernua (Moazzeni 89–26). j inaperturate pollen in O. camptolepis (GZ690). k, l inaperturate pollen in O. stocksii. Scale bars 3 lm (a, b, c, d, e, f, g, j, k), scale bars 1 lm (h, i, l)
I b. exine ornamentation verrucate or scabrate to verrucate. Type II. Tricolpate, radially symmetrical pollen (Phelipanche and Cistanche).
II a. exine ornamentation microreticulate-perforate. Type III. Polymorphic aperture (inaperturate, tricolpate, bi- or trisyncolpate).
123
Pollen morphology and evolution in the genus Orobanche L. s.1. and its allied genera Fig. 7 Box-plot graphic for pollen polar axis in the species of Orobancheae examined. The taxa are sorted by the mean value
III a. exine ornamentation scabrate or verrucate (some members of Orobanche). III b. exine ornamentation microreticulate (some members of Phelipanche). Pollen grains of Orobanche and Diphelypaea are inaperturate and rather asymmetrical, whereas those of Phelipanche and Cistanche are tricolpate and radially symmetrical. Occurrence of tricolpate pollen in Cistanche, Phelipanche and of inaperturate pollen in Orobanche is congruent with results from Abu Sbaih et al. (1994). Minkin and Eshbaugh (1989) investigated some representatives of the New World sections of Orobanche s.l., whose exine ornamentation could be defined as scabrate, microreticulate or verrucate. Microreticulate exine is the most common type in Phelipanche and Cistanche (Figs. 2, 3), while scabrate and verrucate exine ornamentation is common in Orobanche and Diphelypaea (Figs. 4, 5 and 6). The obtained results confirm that pollen characters are not useful for species identification. However, the separation of Phelipanche from Orobanche by Holub (1990) is supported by aperture type and exine ornamentation. Verrucate exine ornamentation and inaperturate pollen type is shared between Orobanche and Diphelypaea, suggesting a close relationship between Orobanche and Diphelypaea. On the other hand, Cistanche has a similar exine pattern and pollen type as Phelipanche. Diphelypaea is included in subtribe Orobanchinae together with Orobanche and Cistanche, while Phelipanche constitutes a separate subtribe Phelipanchinae (Teryokhin et al. 1993, 1997). This view is supported by nuclear ITS (Fig. 9), karyological and genome size data, which indicate that Diphelypaea is closely related to Orobanche (both share a chromosome base number of x = 19), while the Phelipanche and the New World lineages of Orobanche s. l. (all with chromosome base number x = 12) constitute a separate lineage (Schneeweiss et al. 2004a, b; Weiss-Schneeweiss et al. 2006). The phylogenetic position of Cistanche inferred from molecular data is, however, contradicting Teryokhin’s
classification, because rbcL, matK (Young et al. 1999) and ITS-rps2 results (Park et al. 2008) (Fig. 10) indicate a closer relationship of Phelipanche than of Cistanche to Orobanche. Pollen character evolution In phylogenetic research, the presence of tricolpate pollen (or aperture patterns derived from this) is considered as a plesiomorphic trait for eudicots and is widely distributed in most angiosperm families. Furness and Rudall (2004) suggest that optimization of polar aperture character on angiosperm phylogenies indicates an evolutionary transition at the base of the eudicot clade from one aperture to three. In general, more pollen apertures usually offer a selective advantage because it increases the number of prospective germination sites, thus facilitating contact between at least one aperture and the stigmatic surface (Furness and Rudall 2004). Our results and previous studies (Abu Sbaih et al. 1994; Minkin and Eshbaugh 1989; Lu et al. 2007) on this family show that most studied taxa have predominantly tricolpate pollen supporting tricolpate pollen as a plesiomorphic trait in this group. The occurrence of inaperturate pollen in Orobanche and Diphelypaea can be suggested as a synapomorphy. Nowicke (1994) reported that inaperturate pollen occurs in at least 1,500 species in a large group of eudicots. Furness (2007) suggests that fertile inaperturate pollen has evolved numerous times independently throughout eudicots and it may have a functional significance in different groups of the plants like parasitic plants, and our results support this hypothesis. Some of the inaperturate pollen show exine structures, visible under SEM and even light microscopy (Figs. 1, 2, 3 and 4) that resembles a colpus. However, they are thin and found all around the pollen, where exine ornamentation is absent. Abu Sbaih et al. (1994) described these structures as a vestigial colpus. Exine structure is so weak
123
G. Zare et al.
in this part and sometimes causes irregular folding of dry pollen grains. Abu Sbaih et al. (1994) reported this structure in three species of Orobanche; O. alba, O. lutea and O. flava. He suggests that the occurrence of vestigial colpus in this group may indicate an intermediate position between Phelipanche and Orobanche, as this is considered a step from the less advanced tricolpate to the more advanced inaperturate pollen type. Our results show that these structures are found not only in these three species, but also in most Orobanche species particularly in the minor complex taxa. Also, this vestigial colpus is more remarkable in some species such as O. lutea and O. caryophyllacea. Therefore, this character cannot be used for identification of a particular group and treated as intermediate step in evolutionary process (Fig. 11). Reduction of pollen exine and the accompanying loss of apertures may be favored in parasites due to the cost of exine production and could be part of an iterative trend towards morphological reduction in these taxa (Furness 2007). It is suggested that the appearance of vestigial colpi and thin exine site in inaperturate pollen of Orobanche would help to increase the chance for germination of pollen and may be an adaption to complete their life cycle in a short time. Pollen heteromorphism
Fig. 8 Box-plot graphic of pollen polar axis in different populations of the genus Orobanche
123
Whereas most of the studied taxa have homogenous pollen, a group of taxa especially in Orobanche shows pollen heteromorphism. These taxa produce inaperture, tricolpate, bi or trisyncolpate pollen grains in same anther (Fig. 6). All members of the Orobanche section Inflatae (Beck) Tzvel. (except O. stocksii Boiss) and O. anatolica and three taxa from Phelipanche (P. aegyptiaca, P. ramosa and P. schultzii) have heteromorphic pollen grains. Pollen heteromorphism is defined as the production of several pollen grain morphs that differ in aperture number by the same plant (Till-Bottraud et al. 1995). This phenomenon occurs in over 30 % of angiosperm species (Mignot et al. 1994). The main advantage of different pollen morph production in plants is that changes in aperture number may influence parameters of pollen grain fitness (Nadot et al. 2000). A game theory model established by Till-Bottraud et al. (1994) predicts that pollen heteromorphy is an Evolutionary Stable Strategy (ESS) if there is a trade-off between germination speed and life expectancy of the pollen grain, and if pollination conditions vary in an unpredictable way. Natural selection should favor many aperturate quickly germinating pollen grains when growth conditions are suitable and longerlived pollen grains with fewer apertures when conditions are unsuitable for parasitic life. Even if aperture heteromorphism might offer an evolutionary advantage due to
Pollen morphology and evolution in the genus Orobanche L. s.1. and its allied genera
Fig. 9 Box-plot graphic of pollen polar axis in different populations of the genus Phelipanche
group to survive under different ecological conditions and produce rich populations with extensive distribution and a wide range of hosts. As shown by the analyses (Fig. 9), the 3-aperturate pollen morph is the ancestral condition in Orobancheae. Therefore, pollen heteromorphism and the occurrence of 2- and 4-aperturate and inaperturate pollen grains are considered as a derived condition (synapomorphy). Fig. 10 Phylogenetic relationships of Orobanche and Phelipanche with other close relatives such as Boschniakia, Cistanche, Conopholis, Epifagus and Diphelypaea as suggested in the study by Schneeweiss et al. (2004a), using nuclear ITS markers. Phylogenetic tree is constituted with maximum parsimony inference methods and bootstrap support values given in the original studies are indicated for those clades
an increased germination success (Furness and Rudall 2004), it occurs only in limited number of taxa in these genera. Since pollen heteromorphy is often correlated with variation in the sporophytic ploidy level (Bronckers 1963; Erdthman 1969), we looked for sporophyte ploidy level in literature. Variation related to different levels of ploidy occurs in grain size and aperture numbers (Borsch and Wilde 2000). Based on karyological studies conducted by Weiss-Schneeweiss et al. (2006) on the genus Orobanche, chromosome numbers of most of the taxa with polymorph pollen are the same (2n = 2x = 36 for Orobanche and 2n = 2x = 24 for Phelipanche). Pollen heteromorphism is widespread among pest species of Orobanche and Phelipanche, suggesting that it might be induced by atypical ecological circumstances. Also, according to our field experience, these taxa produce rich populations and are widespread. Nadot et al. (2000) suggest that aperture heteromorphism is a good strategy for survival. We suggest that aperture heteromorphism may provide an advantage for this parasite
Pollen size In general, size difference between species is small and consequently, the parameters for polar and equatorial diameters have a low discriminant value within the genus. However, none of the genera or species can be classified by length. In all the taxa, P length shows overlap across species and across genera. In addition, compared with Phelipanche as it is seen in the Box-plot graphic given for P for population and species, there is no correlation between pollen size and classification of these taxa (Fig. 7). However, two genera, Orobanche and Phelipanche, with 37 species, and extensive sampling clearly show that Orobanche compared to Phelipanche has a wider range of variation in size, exine ornamentation and pollen type. And, the genera of Phelipanche have more homogenous pollen grains, compared to Orobanche. Because of the limited number of specimen and of material for the two genera of Cistanche and Diphelypaea, we could not have an exact conclusion about the infrageneric relationship and the variation in these two genera. Further work on this genus still needs to be undertaken, especially on the taxa showing heteromorphic pollen types. Acknowledgments We are grateful to G.M. Schneeweiss (University of Vienna) for his valuable comments and improving the English in the manuscript. We would like to thank Dr. Evren C¸ubukc¸u for his
123
G. Zare et al.
Fig. 11 Phylogenetic relationships in Orobanchaceae inferred from the maximum likelihood analysis of a combined data set plastid rps2 and nuclear ITS sequences (Piednoel et al. 2012). The bootstrap support values given in the original studies are indicated for those clades
help with the scanning electron microscopy at Hacettepe University. The authors would like to thank the Scientific and Technical Research ¨ BI˙TAK-BI˙DEB 2215) for their financial Council of Turkey (TU support to the first author. This research received support from the SYNTHESYS Project which is financed by the European Community Research Infrastructure Action under the FP7 ‘‘Capacities’’ Program for visiting European herbaria.
123
Appendix See Table 2.
Inaperturate
Inaperturate
Kayseri, GZ 592
Kayseri, GZ 593
Kayseri, GZ 594
Ankara, GZ 419 Elazıg˘, GZ 575
O. alba
O. alba
O. alba
O. anatolica
Antalya, GZ 553
Van, FT 1007-2
Van, FT 1009c2
Van, FT 1009a
O. camptolepis
O. caryophyllacea
O. caryophyllacea
O. caryophyllacea
Van, FT 1009b
Antalya, GZ 552
O. camptolepis
O. caryophyllacea
Antalya, GZ 653
Ankara, GZ 690
O. anatolica
O. camptolepis
O. anatolica
Inaperturate
Kayseri, GZ 591
O. alba
Inaperturate
Inaperturate
Inaperturate
Inaperturate
Inaperturate
Inaperturate, Tricolpate, Syncolpate
Inaperturate, Tricolpate, Syncolpate
Inaperturate, Tricolpate, Syncolpate
Inaperturate, Tricolpate, Syncolpate Inaperturate Syncolpate
Syncolpate
Inaperturate
Inaperturate
Inaperturate
Sivas, GZ 499
Sivas, GZ 529
O. alba
Inaperturate
Inaperturate
Inaperturate
Inaperturate
Inaperturate
Inaperturate
O. alba
Kayseri, GZ 491
Kayseri, GZ 494
O. alba
O. alba
O. alba
Sivas, AAD 14855
Erzurum, AAD 14875 Kayseri, GZ 488
O. alba
O. alba
Hakkari, AG 1754
Tunceli, S¸ Y 3245
D. coccinea
D. tourneforti
Inaperturate
Osmaniye, AAD 10686
Diphelypaea coccinea
Tricolpate
Kayseri, M. ¨ ztekin 1491 O
Cictanche salsa
Pollen type
Voucher information
Species
28.97 ± 2.33
32.74 ± 3.30
27.33 ± 1.69
26.97 ± 2.53
21.51 ± 1.76
24.43 ± 0.95
20.70 ± 1.23
30.10 ± 1.72
24.79 ± 1.47
24.20 ± 1.33
26.43 ± 3.84
26.67 ± 2.20
26.50 ± 2.68
26.67 ± 1.99
24.36 ± 1.15
23.94 ± 1.49
25.39 ± 1.43
25.03 ± 1.11
25.20 ± 2.35
25.88 ± 1.76
24.60 ± 1.45
25.35 ± 1.29
24.95 ± 1.39
27.52 ± 1.85
29.1 ± 1.94
P (lm)
28.97 ± 2.33
32.74 ± 3.30
27.33 ± 1.69
26.97 ± 2.53
21.51 ± 1.76
24.43 ± 0.95
21.4 ± 1.13
30.10 ± 1.72
25.03 ± 1.12
24.20 ± 1.33
26.43 ± 3.84
26.67 ± 2.20
26.50 ± 2.68
26.67 ± 1.99
24.36 ± 1.15
23.94 ± 1.49
25.39 ± 1.43
25.03 ± 1.11
25.20 ± 2.35
25.88 ± 1.76
24.60 ± 1.45
25.35 ± 1.29
24.95 ± 1.39
27.52 ± 1.85
26.85 ± 1.69
E (lm)
1
1
1
1
1
1
0.96
1
0.99
1
1
1
1
1
1
1
1
1
1
1
1
1
1
1
1.08
P/E
1.10 ± 0.20
1.30 ± 0.40
1.10 ± 0.20
1.00 ± 0
0.90 ± 0.20
0.96 ± 0.40
1.05 ± 0.16
1.05 ± 0.16
1.02 ± 0.06
1.05 ± 0.15
1.20 ± 0.33
1.30 ± 0.30
1.00 ± 0.22
1.30 ± 0.40
1.00 ± 0
0.90 ± 0.10
1.17 ± 0.32
1.25 ± 0.34
1.22 ± 0.24
1.40 ± 0.30
1.30 ± 0.40
0.9 ± 0.10
0.92 ± 0.10
0.94 ± 0.09
0.95 ± 0.08
Exine thickness (lm)
Table 2 Pollen characteristics of Orobancheae specimens examined in this study and their voucher information
1.00 ± 0
1.00 ± 0
0.90 ± 0.20
1.00 ± 0
0.80 ± 0.20
0.90 ± 0.20
0.96 ± 0.08
0.98 ± 0.06
0.94 ± 0.09
1.00 ± 0
1.00 ± 0
1.00 ± 0
1.00 ± 0
1.00 ± 0
1.00 ± 0.10
0.88 ± 0.10
1.00 ± 0
1.00 ± 0
1.00 ± 0
1.10 ± 0.20
0.90 ± 0.20
1.09 ± 0.16
0.96 ± 0.12
1.1 ± 0.21
1.0 ± 0.1
Intine thickness (lm)
3.80 ± 0.35
4.20 ± 0.63
5.95 ± 1.20
–
–
–
–
4. 40 ± 1.20
–
–
–
–
–
–
–
–
–
–
–
–
–
–
–
–
6.44 ± 2.24
Apocolpium index (lm)
–
–
–
–
14.10 ± 0.67
18.20 ± 0.67
16.30 ± 0.67
–
18.71 ± 1.64
–
–
–
–
–
–
–
–
–
–
–
–
–
–
–
19.1 ± 2.64
Length of colpi (lm)
–
–
–
–
2.90 ± 0.71
4.20 ± 0.60
3.50 ± 0.71
2.20 ± 0.42
3.64 ± 1.28
3.80 ± 1.12
–
–
–
–
–
–
–
–
–
–
–
–
–
–
4.66 ± 0.7
Width of colpi (lm)
Pollen morphology and evolution in the genus Orobanche L. s.1. and its allied genera
123
123
Antalya, AAD 12954
Hatay, GZ 564
Hatay, GZ 626 Antalya, GZ 630
Kayseri, GZ 492
Hatay, AAD 4405
Hatay, GZ 569
Kayseri, GZ 601
O. crenata
O. crenata
O. crenata O. crenata
O. cumana
O. elatior
O. elatior
O. elatior
Aydın, GZ 405 Sinop, GZ 455
Sinop, GZ 463
O. hederea O. hederae
O. hederae
O. hederea
˙Istanbul, GZ 374 ˙Istanbul, GZ 380
O. hadroantha
O. hadroantha
O. hadroantha
O. gracilis
Sinop, AAD 16798 ˙Istanbul, GZ 369 ˙Istanbul, GZ 372
Elazıg˘, GZ 574
O. cernua
O. gracilis
Inaperturate
Iran, Bandar abass, GZ 305
O. cernua
Kayseri, GZ 607
Van, FT 1064
O. cernua
Erzurum, AAD 14886 Sinop, AAD 17338
Hatay, GZ 629
O. caryophyllacea
O. elatior
Inaperturate
Kayseri, GZ 590
O. caryophyllacea
O. fuliginosa
Inaperturate
Elazıg˘, GZ 589
O. caryophyllacea
Inaperturate
Inaperturate Inaperturate
Inaperturate
Inaperturate
Inaperturate
Inaperturate
24.23 ± 1.67
19.26 ± 1.98 25.05 ± 1.66
23.02 ± 1.80
19.37 ± 1.92
23.09 ± 1.71
23.81 ± 1.35
25.63 ± 1.79
25.00 ± 1.35
Inaperturate Inaperturate
24.24 ± 1.38
25.81 ± 2.08
30.25 ± 2.82
26.42 ± 3.57
23.63 ± 1.80
18.29 ± 1.33
23.90 ± 1.17 23.50 ± 1.39
22.36 ± 1.89
25.00 ± 1.21
23.51 ± 1.75
20.30 ± 1.32
22.84 ± 1.43
26.80 ± 1.78
27.76 ± 2.29
31.77 ± 2.33
P (lm)
Inaperturate
Inaperturate
Inaperturate
Inaperturate Tricolpate Syncolpate
Inaperturate Inaperturate
Inaperturate
Inaperturate
Inaperturate, Tricolpat, Syncolpate
Tricolpate, Syncolpate
Inaperturate,
Inaperturate, Tricolpate, Syncolpate
Inaperturate
Inaperturate
Pollen type
Voucher information
Species
Table 2 continued
24.23 ± 1.67
19.26 ± 1.98 25.05 ± 1.66
23.02 ± 1.80
19.37 ± 1.92
23.09 ± 1.71
23.81 ± 1.35
25.63 ± 1.79
25. 00 ± 1.35
24.24 ± 1.38
25.81 ± 2.08
30.25 ± 2.82
26.42 ± 3.57
23.63 ± 1.80
19.20 ± 1.19
23.90 ± 1.17 23.50 ± 1.39
22.36 ± 1.89
25.00 ± 1.21
23.51 ± 1.75
21.87 ± 2.34
22.78 ± 1.62
26.80 ± 1.78
27.76 ± 2.29
31.77 ± 2.33
E (lm)
1
1 1
1
1
1
1
1
1
1
1
1
1
1
0.95
1 1
1
1
1
0.96
0.99
1
1
1
P/E
0.85 ± 0.23
0.78 ± 0.23 0.93 ± 0.27
1.00 ± 0
1.05 ± 0.15
0.85 ± 0.23
1.10 ± 0.20
1.45 ± 0.42
1.00 ± 0
1.20 ± 0.30
1.20 ± 0.20
1.00 ± 0
1.03 ± 0.17
1.20 ± 0.20
0.85 ± 0.15
1.05 ± 0.16 0.96 ± 0.08
1.00 ± 0
1.10 ± 0.20
0.98 ± 0.06
1.10 ± 0.20
1.15 ± 0.24
1.30 ± 0.35
0.91 ± 0.16
0.90 ± 0.20
Exine thickness (lm)
0.90 ± 0.20
0.90 ± 0.20 0.90 ± 0.20
0.98 ± 0.06
0.89 ± 0.16
0.85 ± 0.23
1.10 ± 0.20
0.98 ± 0.06
1.00 ± 0
1.00 ± 0
0.94 ± 0.15
1.00 ± 0
0.98 ± 0.06
0.90 ± 0.20
0.83 ± 014
0.92 ± 0.1 0.90 ± 0.1
0.98 ± 0.06
1.00 ± 0
0.98 ± 0.06
0.98 ± 0.06
1.00 ± 0
1.00 ± 0.1
0.77 ± 0.20
0.90 ± 0.20
Intine thickness (lm)
4.40 ± 1.52
4.10 ± 1.42
4.40 ± 1.43
–
– –
–
–
–
–
–
–
–
–
–
–
–
10.20 ± 2.62
– –
–
–
–
–
–
Apocolpium index (lm)
–
– –
–
–
–
–
–
–
–
–
–
–
–
15.80 ± 1.22
– –
–
–
18.20 ± 0.67
21.10 ± 1.11
16.60 ± 1.07
–
–
–
Length of colpi (lm)
–
– –
–
–
–
–
–
–
–
–
–
–
–
3.40 ± 0.90
– –
–
–
2.90 ± 0.71
2.28 ± 0.22
2.40 ± 0.51
–
–
–
Width of colpi (lm)
G. Zare et al.
Inaperturate
Sinop, AAD 16795 Mersin, GZ 555
Hatay, GZ 565
O. minor
O. minor
Inaperturate Inaperturate
Kars, GZ 466
Kars, GZ 473
Kars, GZ 475
Kars, GZ 476
Kars, GZ 477
O. owerini
O. owerini
O. owerini
O. owerini
O. owerini
Inaperturate
Inaperturate Inaperturate Inaperturate
˙Istanbul, GZ 379 I˙stanbul, GZ 386
O. pubescens
O. pubescens O. pubescens
Aydın, GZ 410
Aydın, GZ 414 ˙Istanbul, GZ 647
O. pubescens
O. pubescens
O. pubescens
Aydın, GZ 401
Aydın, GZ 404
O. pubescens
O. pubescens
O. pubescens
Artvin, GZ 486 ˙Istanbul, GZ 360 ˙Istanbul, GZ 367
O. picridis
Inaperturate
Inaperturate
Inaperturate
Inaperturate
Inaperturate
Inaperturate
Inaperturate
Inaperturate
Kars, GZ 478
Hatay, GZ 566
O. owerini
O. palaestina
Inaperturate
Inaperturate
Inaperturate
Inaperturate
Inaperturate
Kayseri, GZ 596
Van, FT 1061
O. minor
Inaperturate
Inaperturate
Inaperturate
Inaperturate
Inaperturate
Inaperturate
Inaperturate
Inaperturate
Inaperturate
O. minor
O. minor
Kars, GZ 480
Artvin, GZ 484
O. lutea
O. lutea
Kars, GZ 469
Kars, GZ 470
O. lutea
O. lutea
Bitlis, AAD 10994
Kars, S¸ Y 1328
O. kurdica
Malatya, GZ 658 Van, FT 1008
O. kurdica O. kurdica
O. longibracteata
Inaperturate Inaperturate
Antalya, GZ 640
O. hederea
Inaperturate Inaperturate
Denizli, GZ 588
Antalya, GZ 632
O. hederae
Pollen type
O. hederae
Voucher information
Species
Table 2 continued
24.10 ± 1.54
22.05 ± 1.47
24.30 ± 1.43
17.86 ± 1.15
24.24 ± 1.84
23.33 ± 2.09 22.29 ± 1.70
23.81 ± 1.89
24.18 ± 1.20
23.02 ± 1.33
24.48 ± 2.33
25.15 ± 1.49
27.42 ± 1.88
25.45 ± 1.53
24.48 ± 1.37
23.57 ± 1.73
24.36 ± 1.46
23.14 ± 1.34
25.15 ± 2.00
26.48 ± 3.25
23.57 ± 1.11
27.34 ± 33.8
21.68 ± 1.26
24.54 ± 1.51
24.54 ± 1.61
25.64 ± 1.53
24.54 ± 1.32
29.88 ± 2.30
22.24 ± 1.29 26.06 ± 1.71
22.70 ± 1.39
24.30 ± 1.10
25.03 ± 1.55
P (lm)
24.10 ± 1.54
22.05 ± 1.47
24.30 ± 1.43
17.86 ± 1.15
24.24 ± 1.84
23.33 ± 2.09 22.29 ± 1.70
23.81 ± 1.89
24.18 ± 1.20
23.02 ± 1.33
24.48 ± 2.33
25.15 ± 1.49
27.42 ± 1.88
25.45 ± 1.53
24.48 ± 1.37
23.57 ± 1.73
24.36 ± 1.46
23.14 ± 1.34
25.15 ± 2.00
26.48 ± 3.25
23.57 ± 1.11
27.34 ± 33.8
23.15 ± 1.43
24.54 ± 1.51
24.54 ± 1.61
25.64 ± 1.53
24.54 ± 1.32
29.88 ± 2.30
22.24 ± 1.29 26.06 ± 1.71
22.70 ± 1.39
24.30 ± 1.10
25.03 ± 1.55
E (lm)
1
1
1
1
1
1 1
1
1
1
1
1
1
1
1
1
1
1
1
1
1
1
0.9
1
1
1
1
1
1 1
1
1
1
P/E
0.96 ± 0.08
1.08 ± 0.22
1.10 ± 0.20
0.78 ± 0.23
1.20 ± 0.33
1.10 ± 0.20 0.98 ± 0.23
1.03 ± 1.16
0.95 ± 0.27
1.17 ± 0.31
0.90 ± 0.20
1.10 ± 0.20
1.40 ± 0.44
1.30 ± 0.40
1.20 ± 0.33
1.50 ± 0.40
0.89 ± 0.16
1.05 ± 0.15
1.10 ± 0.20
1.20 ± 0.40
1.10 ± 0.20
1.00 ± 0.30
1.00 ± 0.09
1.20 ± 0.33
1.50 ± 0.14
0.98 ± 0.23
1.20 ± 0.25
1.30 ± 0.40
1.05 ± 0.16 1.20 ± 0.20
1.00 ± 0.0
0.98 ± 0.06
1.00 ± 0
Exine thickness (lm)
1.00 ± 0.0
1.05 ± 0.15
1.00 ± 0
0.90 ± 0.20
0.95 ± 0.15
1.00 ± 0 0.93 ± 0.16
1.05 ± 0.15
0.80 ± 0.20
0.95 ± 0.15
0.85 ± 0.23
1.00 ± 00
1.00 ± 0
1.05 ± 0.15
1.10 ± 0.20
1.00 ± 0
1.00 ± 0
0.98 ± 0.06
1.00 ± 0.30
1.00 ± 0
0.90 ± 0.20
0.90 ± 0.20
0.98 ± 0.06
1.00 ± 0
1.00 ± 0
1.00 ± 0
0.96 ± 0.08
1.00 ± 0
0.98 ± 0.06 1.10 ± 0.30
0.92 ± 0.1
0.96 ± 0.08
1.00 ± 0
Intine thickness (lm)
–
–
–
–
–
– –
–
–
–
–
–
–
–
–
–
–
–
–
–
–
–
–
–
–
–
–
– –
–
–
–
Apocolpium index (lm)
–
–
–
–
–
– –
–
–
–
–
–
–
–
–
–
–
–
–
–
–
–
–
–
–
–
–
– –
–
–
–
Length of colpi (lm)
–
–
–
–
–
– –
–
–
–
–
–
–
–
–
–
–
–
–
–
–
–
–
–
–
–
–
– –
–
–
–
Width of colpi (lm)
Pollen morphology and evolution in the genus Orobanche L. s.1. and its allied genera
123
123 Tricolpate Tricolpate
Hatay, GZ 625a Urfa, BM 10745 Elazıg˘, GZ 577
Kars, GZ 467
Van, FT1063
Kahramanmaras¸ AAD8593 Elazıg˘, GZ 578
Alanya, GZ 623
P. aegyptiaca P. aegyptiaca
P. bungeana
P. caesia
P. cilicica
P. coelestis
Tricolpate Tricolpate
Sivas, GZ 495
Sivas, GZ 531
Hatay, GZ 625b I˙stanbul, GZ 351 I˙stanbul, GZ 365 ˙Istanbul, GZ 381
Aydın, GZ 400
Ankara, GT 1776 ˙Istanbul, GZ 370
Alanya, GZ 643
P. mutelii
P. mutelii
P. mutelii
P. nana
P. nowackiana
P. oxyloba
Ankara, GZ 691
Sivas, GZ 497
Sivas, GZ 507
Sivas, GZ 509 Serik, GZ 438
Antalya, GZ 440
P. oxyloba
P. purpurea
P. purpurea
P. purpurea P. ramosa
P. ramosa
P. oxyloba
P. nana
P. nana
P. nana
Sivas, GZ 514 Sinop, GZ 457
P. lavandulacea P. mutelii
Tricolpate
Tricolpate Syncolpate
Tricolpate Tricolpate
Tricolpate
Tricolpate
Tricolpate
Tricolpate
Tricolpate
Tricolpate
Tricolpate
Tricolpate
Tricolpate
Tricolpate
Tricolpate
Tricolpate Tricolpate
Tricolpate
Artvin, GZ 485
Sivas, GZ 532
P. heldereichii
Tricolpate
27.27 ± 1.75
Tricolpate
22.00 ± 1.63
20.89 ± 1.41 20.84 ± 1.52
20.66 ± 1.33
22.23 ± 0.86
22.11 ± 1.07
22.00 ± 1.56
21.44 ± 1.52
24.30 ± 1.56
19.62 ± 0.64
19.99 ± 1.15
19.68 ± 0.90
20.84 ± 1.31
25.30 ± 1.43
23.09 ± 2.30
21.50 ± 1.02
22.35 ± 1.35 19.56 ± 1.20
24.10 ± 1.87
21.68 ± 1.31
23.69 ± 1.86
22.80 ± 1.19
22.00 ± 1.20
24.66 ± 1.39
22.65 ± 1.39
22.60 ± 1.64
25.90 ± 1.66
21.9 ± 1.74 25.03 ± 1.97
27.10 ± 1.95
30. 56 ± 1.95
32.08 ± 2.98
P (lm)
Tricolpate
Tricolpate
Tricolpate
Tricolpate
Tricolpate Syncolpate Tricolpate Syncolpate
Inaperturate
P. heldereichii
Malatya, GZ 654
Sivas, AAD14858
P. coelestis
P. helderechii
P. cilicica
P. arenaria
Malatya, GZ 656
O. stocksii
Inaperturate Inaperturate
Kayseri, GZ 606
Kayseri, GZ 600
O. reticulata
Pollen type
O. reticulata
Voucher information
Species
Table 2 continued
22.54 ± 1.08
22.17 ± 1.23 23.87 ± 1.20
21.75 ± 1.43
23.33 ± 1.19
22.96 ± 1.22
23.9 ± 1.08
20.4 ± 1.19
25.20 ± 1.42
20.84 ± 1.20
22.23 ± 1.67
21.44 ± 1.62
21.14 ± 2.72
25.10 ± 1.28
23.09 ± 2.30
22.20 ± 1.16
22.35 ± 1.35 21.74 ± 1.43
24.10 ± 1.87
23.02 ± 1.57
24.00 ± 1.64
24.2 ± 1.17
24.00 ± 1.10
27.27 ± 1.75
24.66 ± 1.39
23.87 ± 1.21
23.21 ± 1.32
25.82 ± 1.49
22.00 ± 1.8 23.09 ± 1.44
27.10 ± 1.95
30. 56 ± 1.95
32.08 ± 2.98
E (lm)
0.98
0.9 0.9
0.9
0.95
0.96
0.92
1.05
0.96
0.9
0.9
0.92
0.99
1.00
1
0.97
1 0.89
1
0.94
1
0.94
0.91
1
1
0.94
0.97
1
0.99 1.1
1
1
1
P/E
1.00 ± 0
1.00 ± 0 1.00 ± 0
1.00 ± 0.01
1.02 ± 0.06
1.10 ± 0.21
1.00 ± 0.0
1.08 ± 0.22
1.10 ± 0.21
1.10 ± 0.20
1.10 ± 0.20
1.00 ± 0
1.10 ± 0.20
1.05 ± 0.16
0.98 ± 0.06
1.00 ± 0
1.00 ± 0 1.00 ± 0.22
0.93 ± 0.16
1.00 ± 0.09
0.90 ± 0.20
1.05 ± 0.16
1.00 ± 0.0
0.90 ± 0.20
1.00 ± 0.30
1.00 ± 0
1.05 ± 0.15
0.89 ± 0.16
0.90 ± 0.1 1.20 ± 0.24
1.00 ± 0.0
1.20 ± 0.30
0.80 ± 0.30
Exine thickness (lm)
1.00 ± 0
1.00 ± 0.20 0.95 ± 0.15
1.00 ± 0.10
1.00 ± 0.01
1.00 ± 0.0
0.98 ± 0.06
0.98 ± 0.06
1.00 ± 0
0.98 ± 0.06
0.96 ± 0.08
0.96 ± 0.0.8
0.96 ± 0.08
1.00 ± 0.10
0.89 ± 0.16
0.91 ± 016
1.00 ± 0 0.98 ± 0.06
0.86 ± 0.20
0.96 ± 0.08
0.90 ± 0.10
0.96 ± 0.08
1.00 ± 0.10
0.82 ± 0.18
1.02 ± 0.20
1.00 ± 0
0.93 ± 0.15
0.80 ± 0.20
1.10 ± 0.21 1.00 ± 0
0.98 ± 0.06
1.00 ± 0
0.88 ± 0.20
Intine thickness (lm)
10.20 ± 1.82
9.48 ± 1.31 10.45 ± 1.55
10.10 ± 1.18
11.90 ± 1.31
5.40 ± 1.17
5.20 ± 0.92
6.52 ± 1.19
5.10 ± 0.73
6.50 ± 1.02
5.42 ± 1.19
6.32 ± 1.19
7.53 ± 2.02
6.60 ± 0.52
6.08 ± 2.24
8.99 ± 2.18
7.53 ± 1.70 8.75 ± 1.94
5.60 ± 1.10
8.01 ± 1.64
10.0 ± 1.70
4.10 ± 0.74
7.50 ± 0.7
5.60 ± 0.52
10.80 ± 1.31
4.30 ± 1.15
9.48 ± 2.54
5.52 ± 1.20
4.9 ± 0.88 09.18 ± 1.45
–
–
–
Apocolpium index (lm)
16.30 ± 1.11
15.60 ± 3.80 17.30 ± 1.31
17.00 ± 1.3
18.00 ± 1.60
17.10 ± 0.88
18.80 ± 1.48
17.71 ± 1.50
18.80 ± 2.04
17.80 ± 1.21
18.74 ± 1.50
17.74 ± 1.11
18.71 ± 1.55
19.1 ± 0.88
15.00 ± 1.80
17.30 ± 1.31
17.50 ± 1.0 17.50 ± 0.97
15.80 ± 1.98
18.95 ± 1.53
20.00 ± 1.70
16.70 ± 0.67
18.00 ± 1.0
18.2 ± 0.90
17.10 ± 1.53
17.50 ± 1.43
17.70 ± 1.11
17.40 ± 0.47
18.80 ± 0.79 19.67 ± 1.18
–
–
–
Length of colpi (lm)
4.13 ± 1.11
6.10 ± 1.20 5.00 ± 0.90
5.60 ± 1.10
7.05 ± 0.73
3.40 ± 0.52
4.50 ± 0.53
3.52 ± 1.01
7.30 ± 1.25
5.20 ± 1.54
4.60 ± 1.30
7.78 ± 1.82
5.22 ± 1.54
4.70 ± 0.67
5.60 ± 1.10
5.71 ± 1.33
5.42 ± 1.2 6.10 ± 1.20
6.08 ± 1.22
4.40 ± 1.00
7.30 ± 1.50
4.10 ± 0.57
4.70 ± 0.48
6.20 ± 0.67
7.53 ± 1.37
4.20 ± 0.78
6.32 ± 1.19
5.08 ± 1.20
5.60 ± 0.52 5.42 ± 1.2
–
–
–
Width of colpi (lm)
G. Zare et al.
9.48 ± 1.20
20.85 ± 1.32
5.20 ± 0.90
References
Measurements are given as average ± standard deviation P polar axis, E equatorial axis, AAD Ali A. Do¨nmez, FT Fatemeh Taeb, GT Go¨knur Turgut, GZ Golshan Zare
0.96 ± 0.08 1.09 ± 0.23 0.99 25.76 ± 1.41 25.94 ± 1.47 Denizli, GZ 582 P. schultzii
Tricolpate
5.10 ± 0.73 18.00 ± 1.20 10.00 ± 1.10 1.00 ± 0 1.00 ± 0 0.99 22.3 ± 1.32 21.99 ± 1.43 Sivas, GZ 525 P. schultzii
Tricolpate
3.43 ± 0
3.40 ± 0 19.00 ± 1.50
17.30 ± 1.20 10.40 ± 1.52
10.35 ± 1.31 0.69 ± 0.20
0.90 ± 0.20 0.98 ± 0.06
0.83 ± 0.22 1
1 23.57 ± 1.55
27.22 ± 2.18 27.70 ± 2.35
23.57 ± 1.55 Tricolpate Syncolpate
Tricolpate Syncolpate
Mersin, GZ 560
Mersin, GZ 561
P. ramosa
P. ramosa
Voucher information Species
Table 2 continued
Pollen type
P (lm)
E (lm)
P/E
Exine thickness (lm)
Intine thickness (lm)
Apocolpium index (lm)
Length of colpi (lm)
Width of colpi (lm)
Pollen morphology and evolution in the genus Orobanche L. s.1. and its allied genera
Abu Sbaih HA, Keith-lucas DM, Jury SL, Tubaileh AS (1994) Pollen morphology of genus Orobanche L. (Orobanchaceae). Bot J Linn Soc 116:305–313 Beck Von Mannagetta G (1890) Monographie der Gattung Orobanche. Biblioth Bot, Cassel 19:1–275 Beck Von Mannagetta G (1930) Orobanchaceae. In: Engler A (ed) Das Pflanzenreich 96 (IV.261). Wilhelm Engelmann, Germany, pp 1–348 Bennett JR, Mathews S (2006) Phylogeny of the parasitic plant family Orobanchaceae inferred from phytochrome A. Am J Bot 93:1039–1051 Borsch T, Wilde V (2000) Pollen variability within species, populations, and individuals, with particular reference to Nelumbo. In: Harley MM, Morton CM, Blackmore S (eds) Pollen and spores: morphology and biology. Kew R B G, pp 285–299 Bronckers F (1963) Variations polliniques dans une serie d’autopolyplo artificiels d’Arabidopsis thaliana (L.) Heynh. Pollen Spores 5:233–238 Carlo´n L, Go´mez Casares G, Laı´nz M, Moreno Moral G, Sa´nchez ´ , Schneeweiss GM (2005) Ma´s, a propo´sito de algunas Pedraja O Orobanche L. y Phelipanche Pomel (Orobanchaceae) del oeste del Palea´rtico. Doc Jardı´n Bot Atla´nt Gijo´n 3:1–71 Davis PH, Mill RR, Tan K (1988) Flora of Turkey and the East Aegean Islands. Edinburgh University Press, Edinburgh, vol 10. pp 200–201 Erdthman G (1969) Handbook of Palynology. Hafner Publishing Co., New York Furness CA (2007) Why does some pollen lack apertures? A review of inaperturate pollen in eudicots. Bot J Linn Soc 155:29–48 Furness CA, Rudall PJ (2004) Pollen aperture evolution – a crucial factor for eudicot success? Trends Plant Sci 9:154–158 Gilli A (1971) Die Orobanchen der Tu¨rkei. Feddes Repert 82(6):381–406 Gilli A (1982) Orobanche L.. In: Davis PH (ed) Flora of Turkey and The East Aegean Islands, vol 7. pp 1–23 Hesse M, Halbritter H, Zetter R, Weber M, Buchner R, Frosch– Radivo A, Ulrich S (2009) Pollen Terminology. An illustrated Handbook. Springer, Vienna. http://www.paldat.org/paldatTerminology-large.pdf Holub J (1977) New names in Phanerogamae 6. Folia Geobot Phytotax 12:417–432 Holub J (1990) Some taxonomic and nomenclatural changes within Orobanche s. l. (Orobanchaceae). Preslia 62:193–198 Lu L, Wang H, Blackmore S, Li DZ, Dong LN (2007) Pollen morphology of the tribe Rhinantheae (Orobanchaceae) and its systematic significances. Pl Syst Evol 268:177–198 Mignot A, Hoss C, Dajoz I, Leuret C, Henry JP, Dreuillaux JM, Heberle-Bors E, Till-Bottraud I (1994) Pollen aperture polymorphism in the angiosperms: importance, possible causes and consequences. Acta Bot Gallica 141:109–122 Minkin JP, Eshbaugh WH (1989) Pollen morphology of the Orobanchaceae and Rhinanthoid Scrophulariaceae. Grana 28:1–18 Nadot S, Ballard HE Jr, Creach JB, Dajoz I (2000) The evolution of pollen heteromorphism in Viola: a phylogenetic approach. Pl Syst Evol 223:155–171 Park JM, Manena JF, Colwell AE, Schneeweiss GM (2008) A plastid gene phylogeny of the non-photosynthetic parasitic Orobanche L. (Orobanchaceae) and related genera. J Plant Res 121:365–376 Piednoe¨l M, Aberer AJ, Schneeweiss GM, Macas J, Novak P, Gundlach H, Temsch EM, Renner SS (2012) Next-generation
123
G. Zare et al. sequencing reveals the impact of repetitive DNA in phylogenetically closely related genomes of Orobanchaceae. Mol Biol Evol 29(11):3601–3611. doi:10.1093/molbev/mss168 Schneeweiss GM, Colwell A, Park JM, Jang CG, Stuessy TF (2004a) Phylogeny of holoparasitic Orobanche (Orobanchaceae) inferred from nuclear ITS-sequences. Mol Phylogent Evol 30:465–478 Schneeweiss GM, Palomeque T, Colwell AE, Schneeweiss HW (2004b) b) Chromosome numbers and karyotype evolution in holoparasitic Orobanche L. (Orobanchaceae) and related genera. Am J Bot 91(3):439–448 Shavvon RSh, Mehrvarz ShS (2010) Pollen and seed morphology of the genus Cistanche (Orobanchaceae) in Iran. Biologia 65(4):615–620 Sojak J (1972) Nomenklatoricke poznamky (Phanerogammae). Casopis Narodniho Pers. Proc Nat Acad Sci India 9:58–68 Teryokhin ES, Schibakina GB, Serafimovitsch NB, Kravtzova TI (1993) Opredelitel Sarasychovych Flory SSSR. ‘‘Nauka’’ Sankt Petersburg, p 123 Till-Bottraud I, Venable DL, Dajoz I, Gouyon P (1994) Selection on pollen morphology: a game theory model. Am Nat 144:395–411
123
Till-Bottraud I, Mignot A, De Paepe R, Dajoz I (1995) Pollen heteromorphism in Nicotiana tabacum (Solanaceae). Am J Bot 82:1040–1048 Uhlich H, Pusch J, Barthel KJ (1995) Die Sommerwurzarten Europas Weiss-Schneeweiss H, Greilhuber J, Schneeweiss GM (2006) Genome size evolution in holoparasitic Orobanche (Orobanchaceae) and related genera. Am J Bot 93:148–156 Wodehouse RP (1935) Pollen Grains. Mc Graw-Hill, New York Young ND, Steiner KE, dePamphilis CW (1999) The evolution of parasitism in Scrophulariaceae/Orobanchaceae: plastid gene sequences refute an evolutionary transition series. Ann Miss Bot Gard 86:876–893 Zare G (2012) A taxonomic revision of the genus Orobanche L. (Orobanchaceae) in Turkey, it’s relation with the Iranian taxa and their molecular phylogeny. Ankara, Bot Dept Hacettepe Univ. PhD thesis Zare G, Do¨nmez AA (2013) Two new records of the genus Orobanche (Orobanchaceae) from Turkey. Turk J Bot 37:597–603 Zare G, Do¨nmez AA, Koca AD (2010) A new record for the flora of Turkey: Orobanche palaestina Reut. (Orobanchaceae). Hacettepe J Biol Chem 38(2):149–154
