Acta Bot. Croat. 79 (1), 59–67, 2020 CODEN: ABCRA 25 DOI: 10.37427/botcro-2020-006 ISSN 0365-0588 eISSN 1847-8476 Morphology, anatomy, palynology and achene micromorphology of Bellis L. (Asteraceae) species from Turkey Faruk Karahan* Department of Biology, Faculty of Science and Arts, Hatay Mustafa Kemal University, 31040 Hatay, Turkey Abstract – In the present study, the morphological characters, root, stem and leaf anatomy, pollen and achene micromorphology of Bellis L. species (Bellis annua L., B. perennis L. and B. sylvestris Cirillo) distributed in Turkey have been investigated on light and scanning electron microscope. Palynological analysis showed that pollen characters were found as small to medium size, isopolar, radially symmetrical, oblate-spheroidal and prolate- spheroidal, tricolporate and echinate-perforate ornamentation in the three species. Achene characters were found dark brown to yellow in colour, often cylindrical, compressed, with thickened margin, obovate orobovoid shaped, pappus absent and the coat ornamentations are rectangular with short hairs on the surface. As a result of this study, leaf morphology and some pollen characteristics such as pollen size, shape, perforation and distance be- tween spines were demonstrated to be different among the Bellis species. Keywords: Bellis, common daisy, Compositae, taxonomy, SEM Introduction The genus Bellis L. (Asteraceae) has been included in the Fiz et al. (2002) studied the phylogenetic relationships subtribe Bellidinae Willk. (tribe Astereae Cass.) along with between Bellis and the closely related genera (Bellidastrum 117 other genera representing more than 3000 annual or Scop, Bellium L. and Rhynchospermum Lindl.) and evolu- perennial taxa (Bremer 1994). It is native to western, cen- tion of their morphological characters. Kavalcıoğlu et al. tral and northern Europe, Cyprus, western Syria and Azer- (2010) did a comparative RAPD analysis and pollen struc- baijan but is commonly found as an invasive plant in North ture studies of B. perennis from Rize and Antalya cities of America (Webb 1976). In Turkey, according to the Flora of Turkey. Bozdağ et al. (2011) published data on the chromo- Turkey and the East Aegean Islands (Grierson 1975) and the some number and morphology of Bellis species of Turkey. last checklist of Turkish Flora (Güner et al. 2012), the genus The chromosome number was determined in B. sylvestris, B. is represented by three species: Bellis annua L., B. perennis L. perennis and B. annua as 2n = 36, 18 and 18, respectively. The and B. sylvestris Cirillo. It is known as „akbubeçlik, nineotu, previous studies reported that pollen and seed micromor- çayır papatyası, koyungözü, koyun çiçeği and yoğurt otu” in phologies are efficient in the systematics of the Asteraceae Anatolia (Baytop 1994, Ekim 2012). family (Moore et al. 1991, Shabestari et al. 2013, Frangiote- The genus Bellis (common daisy) is known as a tradition- Pallone and De Souza 2014). Akyalçın et al. (2011) studied al wound herb (Ai-Douri and Al-Essa 2010) and it was used the pollen morphology of six Achillea L. sect. Achillea (As- for the treatment of bruises, broken bones, and wounds by teraceae) species from Turkey. Their results show that pol- Crusaders in the Middle Ages (Mitich 1997). It has also been len grains of Achillea species were oblate-spheroidal, prolate- used in traditional folk medicine for the treatment of sore spheroidal, subprolate and generally tricolporate, though at throat (Uysal et al. 2010), headache, stomach ache, common times tetracolporate or even pentacolporate. The structure of cold, eczema, wound healing, diarrhea, peptic ulcer, gastri- the exine is double tectate and mean exine thickness varied tis, rheumatism, asthma, hemorrhoids and as a vulnerary, from 3 to 8.5 μm. Özler et al. (2009) investigated the pollen expectorant and laxative (Kavalcioğlu et al. 2010, Karakaş morphology of 29 taxa of the genera Centaurea L., Psephellus et al. 2012, Melikoğlu 2015). Cass. and Cyanus Miller from Turkey. They reported that * Corresponding author e-mail: [email protected] ACTA BOT. CROAT. 79 (1), 2020 59 KARAHAN F pollen grains were tricolporate, rarely tetracolporate, isopo- Specimens were dried according to standard herbari- lar, radially symmetrical, subprolate, spheroidal-subprolate, um techniques and are deposited in the Herbarium of Ha- operculate, tectum perforate, with a microechinate and sca- tay Mustafa Kemal University, Hatay, Turkey (HMKU). brate exine surface. Joujeh et al. (2019) conducted a palyno- The botanical identification was based on the Flora of Tur- logical study on Centaurea iberica Trev. ex Spreng, C. virgata key and the East Aegean Islands (Grierson 1975). In addi- Lam., C. verutum L., C. hyalolepis Boiss., C. solstitialis L., C. tion, herbarium specimens from the Royal Botanic Garden ammocyanus Boiss. from Syria. They reported that pollen (KEW), Royal Botanic Garden Edinburgh, ANK (Herbari- grains of the studied species were similar in shape, size, ap- um Turcicum), AEF (Ankara University Faculty of Pharma- ertures characters, polarization and symmetry, while they cy Herbaryum), GAZI (Gazi University Herbarium), HUB differed in color, exine ornamentation pattern and density of (Hacettepe University Herbarium), KNYA (Selçuk Univer- the spines distributed on the exine surface and in quantita- sity Herbarium) and OMUB (Ondokuz Mayıs University tive measurements. However, studies about pollen and seed Herbarium) herbaria were checked. Synonyms of taxa fol- morphology of Bellis species from Turkey and near related low WFO (2019). countries are limited. The plant samples were preserved in 70% ethanol for an- The aim of the present study is to give a detailed descrip- atomical analysis. Transverse sections of root, stem and leaf tion of the anatomical features of root, stem and leaf, as well samples were prepared. Slides were observed with a Zeiss as of the achene and pollen morphology of Bellis species dis- Axiolab A1 light microscope. For anatomical description, tributed in Turkey, and evaluate the systematic significance. the terminology of Metcalfe and Chalk (1957) was used. To the best of our knowledge this is the first report on all of Pollen grains were obtained from mature anthers of dried the morphological, anatomical and palynological characters flowers and measured under a light microscope from non- of Bellis species growing in Turkey. acetolyzed samples after preparation applying the Wode- house (1935) method. The measurements of the polar axis Materials and methods (P), the equatorial diameter (E), the colpus length (clg), the The materials of this study includes 22 specimens be- ornamentation, the number of apertures (An), the aperture longing to 3 populations of B. annua, 102 specimens be- type, the spine length (spg), the spine width (spt), the ex- longing to 15 populations of B. perennis and 32 specimens ine (e) and the intine (i) thickness for 20 pollen grains were belonging to 4 populations of B. sylvestris that were collected conducted under a Zeiss Axiolab A1 light microscope. In from 22 different localities in Turkey during their flowering the course of scanning electron microscopy studies, mature and fruiting time, between 2014 and 2016 (Tab. 1). The re- achenes and pollen grains were mounted using double-sid- search area is given in Fig. 1 according to Davis’s Grid sys- ed tape on the stubs and coated with platinum-palladium in tem (Davis 1965-1985). a Quorum Q150R EM sputter coater. They were examined Tab. 1. Localities, habitats and sample codes of the studied Bellis L. species in Turkey. Abbreviations: NP – Number of population, NS – Number of specimens, FK – Faruk Karahan. Altitude Collector NP NS Taxa Square, location and habitat Co-ordinates (m a.s.l.) number 1 5 B. annua L. A4: Karabük: Ovacık, Boyalı village, grassland 623 41°07'01.5"N 32°49'25.6"E FK 1210 2 6 B1: Manisa: Beydere village, wet grassland 210 38°40'53.7"N 27°14'54.6"E FK 1218 3 11 C6: Hatay: Kırıkhan, Gölbaşı village, grassland 180 36°30'13.0"N 36°28'28.2"E FK 1102 4 6 B. perennis L. A1(E): Kırklareli: Armağan Dam, forest 448 41°54'14.9"N 27°25'40.2"E FK 1124 5 7 A2: Kocaeli: Karamürsel, Senaiye village, forest 432 40°37'41.7"N 29°41'03.7"E FK 1132 6 5 A3: Bolu: Mengen, Demirciler village, grassland 1450 40°57'23.0"N 32°05'42.2"E FK 1138 7 6 A4: Kastamonu: Azdavay, Kart mountain, forest clearings 1227 41°45'02.9"N 33°17'58.9"E FK 1204 8 7 A7: Giresun: Şebinkarahisar, Tamzara village, meadow 1332 40°19'42.7"N 38°26'07.6"E FK 1103 9 5 B6: Sivas: Suşehri, Sökün village, wet slopes 1150 40°17'07.0"N 38°10'33.2"E FK 1105 10 10 B6: Kayseri: Pınarbaşı, roadside 1520 38°43'13.9"N 36°23'10.4"E FK 1104 11 5 B7: Elazığ: Altınkuşak village, wet grasslands 982 38°47'50.9"N 39°05'33.2"E FK 1321 12 6 C3: Isparta: Yenişarbademli, wet grassland 1147 37°42'21.4"N 31°23'09.3"E FK 1217 13 6 C4: Konya: Tatköy near,wet grasslands 1449 37°56'46.7"N 32°21'29.0"E FK 1189 14 7 C5: Adana: Pozantı, Beypınarı, moist slopes 1532 38°05'53.6"N 36°14'07.9"E FK 1139 15 8 C6: Hatay: Antakya Castle, wet grassland 440 36°13'08.8"N 36°10'48.0"E FK 1106 16 8 C6: Hatay: Kırıkhan, Çataltepe village, grassland 128 36°31'03.8"N 36°25'43.0"E FK 1108 17 6 C6:Osmaniye: Düziçi, Haruniye Castle, grassland 485 37°16'07.0"N 36°29'14.4"E FK 1110 18 8 C7: Şanlıurfa: Birecik, Zeytinbahçe near, grassland 402 36°55'17.5"N 38°01'40.6"E FK 1249 19 8 B.