Abstract: There are 4 species belong to Ankyropetalum Fenzl (Caryophyllaceae) genus and three of them (A. reuteri Boiss. and Hausskn, A. arsusianum Kotschy ex Boiss. and A. gypsophiloides Fenzl) are distributed in Turkey. There are doubts about taxonomical studies depending on only morphological characteristics. This study has been made to put forth that palinological studies also contribute taxonomical studies. Pollen morphology of the three species belong to Ankyropetalum Fenzl (Caryophyllaceae) genus distributed in Turkey examined with ray microscope and electron microscope in this study. Results evaluated according to Duncan’s multiple range test using SPSS statistic program. Pollen’s polar and ecvatoral seeming photographs were taken in preparates. Morphology of pollens examined with 50 repetition for each taxon and morphological assessments were made. The common trait of pollens can be summarized as they are circular, oblate and prolate spheroidal, perporate (pore numbers ranged between 20-33), operculum is granulated, annulus is distinct, the form of pollens are tectat. Definition of pollens are given for each taxon, diagnostic specifications recognized as important are used for making diagnosis key. The difference between species are as below: A. arsusianum’s pollen shape is oblate-spheroidal, type of pollen is periporate, pore numbers are between 23-33, form of pollen is tectat, ornamentation is perforate. A. reuteri’s pollen shape is prolate-spheroidal, type of pollen is periporate, pore numbers are between 20-33, form of pollen is tectat, ornamentation is from perforate to eureticulate A. gypsophiloides pollen shape is oblate-spheroidal, type of pollen is periporate, pore numbers are between 21-30, form of pollen is tectat, ornamentation is perforate.
INTRODUCTION
Caryophyllaceae is big family in respect to genus and species number and important for Türkiye’s economy (Guner et al., 2000; Secmen et al., 2004; Korkmaz and Özçelik, 2011; Özçelik and Yıldırım, 2011). Ankyropetalum Fenzl genus has three species which names are A. reuteri, A. arsusianum and A. gypsophiloides, in Türkiye. One of them is endemic and the others are narrow occurrence (Muca, 2009; Özçelik and Muca, 2010; Çelik et al., 2012). In respect of Ankyropetalum species expansion is usually in southwestern Asia (Afifi and Abu-Irmaileh, 2000). These taxa is which region intersect Irano-Turanian and Mediterranean, grown naturally in small region of Türkiye’s southerneast (Boissier, 1867; Huber-Morath, 1967; Özçelik and Muca, 2010). Outside of Türkiye Ankyropetalum species spread Syria (Mouterde and Premier, 1966), Egypt (Tackholm, 1974), Iran, Lebanon, Sinai Penunsula (Huber-Morath, 1967) and Irak (Mouterde and Premier, 1966).
Now-a-days only morphological characters are’nt enough for taxonomic studies. Especially description isn’t impassable for close genus and species. Anatomy and palynology seems the most close science to morphology science. Ankyropetalum genus has a lot of taxonomic problems and because of these problems Ankyropetalum and Gypsophila genera are confuse frequency each other. There is no study about Ankyropetalum species’ pollens in Türkiye. In our study we inspected species of Ankyropetalum genus’ pollens ornamentation and general appearance with electron microscope (SEM); pollen morphologies with light microscope and determined differences of species. Our studies which about Gypsophila taxa in Türkiye, are going on. When our study has been completed, these close genara could expect to distinguish palynologically.
| Table 1: | Comparing Palynological Characters of Ankyropetalum species’ in Türkiye |
| Data are available in range and Mean±SD | |
MATERIALS AND METHODS
Pollens were prepared up to Wodehouse (1935)’s method. A. gypsophiloides and A. reuteri’s herbarium examples and A. arsusianum’s fresh flowers were used for pollen preparation (Muca, 2009). Light microscope was used for morphological study. There were taken pollens polar and equatoral aspects photos from light microscope. Pollen morphology studies were done 50 repeated process for each taxon. Pollens pores were tallied up to McAndrews and Swanson (1967).
Pollens preparated up to Yoshinori et al. (1991) for inspection ornamentation and general appearance for Scanning Electron Microscope (SEM). The anther parts of the stamens were kept carnoy fixative for 48 h and then pollens were cleaned on microscope slide with 70% alcohol. Pollen grains upper surfaces were covered and scanned on electron microscope after evaporated alcohol and then were taken photos. These photos show pollens ornamentation, pores and general aspect. SEM studies were completed at the university of Mediterranean in medicine faculty’s unit of temga.
The results of this study were interpreted according to Duncan test using SPSS statistic programme (Table 1). Pollen descriptions were given each taxa and diagnostic characters were used for doing diagnosis key.
RESULTS
Ankyropetalum genus has 3 taxa which are A. arsusianum, A. reuteri, A. gypsophiloides in Türkiye. Examples were collected from 10 different localities. Species spread southerneast of our country (Fig. 1).
Ankyropetalum arsusianum Ky, Pl exs Fl. Or. 1: 533 (1867):
| • | Type: C5 Hatay: Amanos Mountain, Arsus locality |
| • | Fl: 6-7 |
| • | Spreading: Türkiye (Osmaniye-Adana, Hatay) and Syria. Although it is not endemic in Türkiye known to collect type narrow place. It is Iran-Turanian element |
| • | Inspected Localities and Collections: C6: Osmaniye: Adana-Gaziantep highway, from Adana 70 km (N: 40°99'014, E: 36°760'859), appears on the surface volcanic rock slopes, 50-100 m, 11.06.2009, Belkıs 04 (Fig. 2) |
A. arsusianum’s pollen features: Pollen shape oblate-spheroidal (A/B = 0,99). Pollens general aspect is circular. A-axis 39.55, 38-41±0.9 μm, B-axis 38-41 μm, 39.6±0.9. Pollen type periporate, pore No. 23-33, 29.1±3,06 μm. Pore height (Plg) 7-10 μm, 7,85±1,04; pore width (Plt) 7-10, 7,85±1.04 μm, cycloid. The distance between the two pore (Pl-Pl) 13-15 μm, 13.75±0.79 μm. Operculum (Op) with granule; granules are distinctive. Annulus (Anl) is distinctive, 1-2 μm, 1, 10±0.26 μm. Exine thicknesses 1-2 μm, 1,5±0,5 μm. Intine thicknesses 1 μm. Pollen structure is tectat, ornamentation is reticulate. Granules are distinctive on exine surfaces (Fig. 3, 4).
A. reuteri Boiss. and Hausskn. (Fl. Or., 1: 533(1867):
| • | Type: C6 Kahramanmaraş, Ahir Daği |
| • | Fl: 6-7 |
| • | Spreading: It is endemic for Antitoros’ in Türkiye. It is known a few collecting type in narrow place. It spreads Kahramanmaraş, Osmaniye and Adiyaman. It seems that widely spread but It is small area which is spreading in different cities. But this area may be more than 50 km diameter. Iran-Turanian element |
| Fig. 1: | Taxa of Ankyropetalum genus’ spreading map |
| Fig. 2: | A. arsusianum’s overview of habitat |
| • | Inspected localities and collections. |
| • | C6: Osmaniye-Kahramanmaraş road, near Kahramanmaraş city border, to Türkoğlu 20 km, to Kahramanmaraş 42 km, (N: 41°21'318, E: 37°30'2429), step, 500-550 m., 06.06.2009, Belkıs 01 |
| • | C6: Adıyaman, between Gölbaşı-Erkenek (Malatya), after 14 km from Gölbaşı., after±2 km Göksu Bridge, Vadi Restaurant, near Çavuşoğlu Service Area (N: 41°91'307, E: 37°39'0563), steppe, volcanic and with erosion hills, 850-900 m., 6.6.2009, Belkıs 02 (Fig. 5) |
| Fig. 3(a-b): | A. arsusianum’s pollen morphology’s aspect on light microscope (a) Optic section and (b) Ornamentation (Horizontally bar for each photo: 10 μm) |
| Fig. 4(a-b): | A. arsusianum’s pollen morphology on SEM (a) Ornamentation and pore aspect and (b) General aspect |
| Fig. 5: | A. reuteri’s overview of habitat |
| • | C6:Kahramanmaraş-Türkoğlu, Çakıroğlu village, there were much more pasturaged steppe areas, 500-600 m., 27.07.2000, Özçelik 8552. |
A. reuteri’s Pollen features: Pollen shape prolate-spheroidal (A/B = 1.02). Pollens general aspect is circular. A axis 38-42 μm, 39.65±1.35 μm. B-axis 32-43 μm. 38.7±2.4. Pollen type periporate, pore number 20-33, 26.7±4.07 μm. Pore height (Plg) 6-9 μm. 8.05±0.94. pore width (Plt) 6-9 μm, 7.85±0.99 μm. cycloid. The distance between the two pore (Pl-Pl) 12-16 μm. 14.25±1.16 μm. Operculum (Op) with granόle and granules are distinctive. Annulus (Anl) is distinctive. 1-1.5 μm, 1.02±0.11 μm. Exine thicknesses 1-2 μm, 1.5±0.5. Intine thicknesses 1-2 μm. 1±0. Pollen structure is tectate. ornamentation perforate to eureticule. Granules are distinctive on exine surfaces (Fig. 6, 7).
A. gypsophiloides Fenzl, (Bot. Zeitschr., 1843: 393 (1843):
| • | Syn: Ankyropetalum coelesyriacum Boiss.; Diagn. Ser. 1(8): 59 (1849). Gypsophila coelesyriaca (Boiss.) |
| Fig. 6(a-b): | A. reuteri’s pollen morphology’s aspect on light microscope (Horizontal bar for each photo: 10 mm) |
| Fig. 7(a-b): | A. reuteri’s pollen morphology on SEM (a) Ornamentation and pore aspect and (b) General aspect |
| Fig. 8: | A. gypsophiloide’s overview of habitat |
| Fig. 9(a-b): | A. gypsophiloides’s pollen morphology’s aspect on light microscope. (a) Optic section and (b) Ornamentation (Horizontally bar for each photo: 10 μm) |
Williams; Jown Bot.(London), 27:322 (1889), Gypsophila subaphylla Rech.; Bot. Jahrb., 75: 355(1952), Gypsophila gypsophiloides (Fenzl) Blakelock; Kew Bull., 2: 193(1957), Gypsophila gypsophiloides (Fenzl) Blakel.; Nouvelle Flore du Liban et de la Syrie, 517(1966):
| • | Type: C8 Mardin, Binibil |
| • | Fl: 6-7 |
| • | Spreading: Türkiye (Southerneast Anatolian), Syria, Iranian, Egypt, Lebanon, Palestinian, Sinai Penunsula. Iran-Turanian’s (Sahara Arabian) Element |
Inspected localities and collections:
| • | B8: Diyarbakır-Silvan (Şanlıurfa) road, to Silvan 50 km, steppe, 800-850 m., 28.07.2000, Özçelik 8563 |
| • | C7: Şanlıurfa City Center, to Adıyaman, Servis Area in the forest in sanliurfa (N: 41°24'195, E: 37°48'4067), near Pinus brutia forest, volkanic land, 850-900 m, 09.06.2009, Belkıs 03 (Fig. 8) |
| • | C8: Siirt-Kurtalan, Gökdoğan (Cimzark) village (N: 38° 08'057, E: 41°18'005), near of Garzan Stream, 800 m., 12.07.1991, Özçelik 2504 |
| • | C9: Siirt-Pervari road, to Pervari 50 km, steppe, 600 m, 15.07.1991, Özçelik 2503, 2555 |
| • | C9: Siirt-Pervari road (N: 37°52'720, E: 41°53'362), near Botan stream, 1000 m., Agust 1991, Özçelik 2564 |
A. gypsophiloides’s pollen features: Pollen shape oblate-spheroidal (a/b = 0.95). Pollens general aspect is circular. A axis 32-43 μm. 37.75±2.9 μM. B-axis 37-42, 39.55±1.5 μM. Pollen type periporate. pore number 21-30. 25.8±2.59. Pore height (Plg) 6-9 μm. 7.15±0.99 μm. pore width (Plt) 6-9 μm. 7.15±0.93 μM cycloid. The distance between the two pore (Pl-Pl) 10-14 μm. 11.75±1.25 μm. Operculum (Op) with granόle and granules are distinctive. Annulus (Anl) distinctive 1-1.5 μm. 1.15±0.23 μm. Exine thicknesses 1-2 μm, 1.45±0.5 μm. Intine thicknesses 1 μm. Pollen structure tectate. ornamentation perforate. Granules are distinctive on exine surfaces (Fig. 9, 10).
| Fig. 10(a-b): | A. gypsophiloides’ s pollen morphology on SEM (a) Ornamentation and pore aspect and (b) General aspect |
DISCUSSION
Erdtman (1943, 1952), Erdman (1969) had searched Caryophyllaceae family pollens on electron microscope, he used data about these pollens and he brought solutions for phylogenetic and systematical problems (Romanova, 1992). Genara of Caryophyllaceae of pollen morphologies were studied a lot of researchers. Pollens which have a lot of pore is characteristic for family. Spergula and Spergularia genus’ are exception genus’ about this subject. The smallest pollen grain’s shape is elliptical-square in Herniaria (Illecebraceae) genus (11- 16,5 μm) and the biggest pollen grains are seemed in Agrostemma githago L. (55-65 μm) taxa (Romanova, 1992). Furthermore 38 taxa of 12 genus of Caryophyllaceae family has been researched (Yıldız, 2001).
Palynology which conduce to angiosperm systematic and phylogeny, is important dicipline. Pollen characters has important place for defining taxonomical categories and separating taxa. Pollen characters is so important for Takhtajan and Cronquist’s angiosperms classification system; pollen morphologies’ similarity and differences had put forth relationship between taxa (Walker and Doyle, 1975; Yıldız, 2001).
Barkoudah (1962) had done revision study about Gypsophila, Bolanthus, Ankyropetalum and Phryna genara. In this study, he had determined that Gypsophila pollens periporate shape.
He had found that frequently 12 (change between 9-16) pore numbers and their size are changing between 16-35.5 μm (median 22 μm). These knowledge are parallel with our results and this explains that relationship between Ankyropetalum genus and Gypsophila Bolanthus, Phyrana genara. It had (Huber-Morath, 1967) which diagnosis key was done with classical taxonomy, emphasized clearly this similarity.
The species of Ankyropetalum’s pollens structure which spread in Türkiye have same circular shape and more pore numbers as the other family members. Generally pollen size is 39 μm (mean value) and pore numbers are 27(mean value). We can see their spines when we investigate ornamentations. Exine and intines are thin like Gypsophila pollens (Ataşlar, 2009).
Up to our results, Ankyropetalum species that spread in Türkiye can distinguish with following diagnosis key:
| 1. | Pollen shape oblate-spheroidal, ornamentation perforat; number of grain on the pores 7-12 | |
| 2. | Pollens length/breadth rate (a/b) 0,95; pore number 21-30;….………gypsophiloides | |
| 2. | Pollens length/breadth rate (a/b) 0.99; pore number 23-33:…………....arsusianum | |
| 2. | Pollen shape prolate-spheroidal, ornamentation perforate to eureticulate; number of grain on the pores 8-14…..…………………………………………………………………………….reuteri | |
If A. colesyriacum add to this diagnostic key, all species of this genus’ pollens will be examine on the world and bring palynological features to taxonomy. Gypsophila species’ of Türkiye have examine by us. When this study completed, these close relative genus’ (Ankyropetalum and Gypsophila) will distinguish each other with clear characters.
Skvarla and Novicke (1976) had done a research about Silene and Dianthus genus’. They had found Silene noctiflora L. and Dianthus barbatus L. species’ pollens that have pantoporate, thin tectum, sharp spines structures. According to our studies, Ankyropetalum pollens spines are circular or near to circular on apical part but never sharp. This feature shows that Ankyropetalum genus have different feature and they are far relationship from Silene and Dianthus genus’.
Ghazanfar (1984) had investigated on Silene genus’ pollen morphologies belonging to Siphonomorpha Otth. and Auriculatae Boiss. Sections. He had found with his study pollens that have pantoporate and spheroid structure, pollens diameters are bigger (30-40 μm) and they have various pore number. Pollens of Ankyropetalum species’ diameters are changing between 32-41 μm and pore number are changing 20-33. Pollen diameters are big as pollens of Silene genus and it is similar to Silene genus with this feature.
Taia (1994) had investigated some Caryophyllaceae examples pollens spreading in Egypt. In Taia’s study that he investigated 67 species in 21 genus and he had reported pollen size, number of aparture and thinness of exine. According to this report, he was done 4 pollen groups (Pantoporate, Trizonocolpate, Pancolpate, Pancolpate and Mix aparture). He had determined 9 type pollen and 4 different pollen shape (spheroidal, subprolate, prolate, oblate). He had determined to similarity of Arenaria and Minuartia genus’ pollens. According to our results, member of Ankyropetalum genus’ pollens aren’t congruity with these groups.
When studies had investigated, almost all genus’ pollen examples had been studied beside Ankyropetalum genus. General feature of family that pollens are circular and have pores. Pollen sizes, ornamentations, pore numbers and diameter of pores are changing each other. Some researchers have separated to different types Caryophyllaceae pollens. Ankyropetalum genus’ palynological features were investigated firstly and by us, results were commented for taxonomic aim.
ACKNOWLEDGMENT
This study is part which name is Revision of Gypsophila L. and Ankyropetalum Fenzl (Caryophyllaceae) Genara and Determination of Saponin Level (Its number is TBAG 107T147 and supported by Tübitak. We thank to presidency of Tübitak and project group with sincerity.
We thank to Asist Prof. Dr. Ali Ince for his helps about measuring pollens size and comment to results and thank to Prof. Dr. Necdet Demir (Mediterranean University, Head of Department of Histology and Embriyology) and Hakan Er (Responsible of Scanning Electron Microscope Unit of Tagem Department) for taking photos of pollens on Scanning Electron Microscope.
We thank to Ahmet Erçin and his family (from Siirt, Kurtalan-Village of Gökdoğan) for interest and support about our study.