Acta Botanica 1-2015 - za web.indd ACTA BOT. CROAT. 74 (1), 2015 31 Acta Bot. Croat. 74 (1), 31–41, 2015 CODEN: ABCRA 25 ISSN 0365-0588 eISSN 1847-8476 Morphological and anatomical features of seeds of Turkish Romulea taxa (Iridaceae) and their taxonomic signifi cance MEHMET CENGİZ KARAİSMAİLOĞLU Istanbul University, Faculty of Science, Department of Biology, 034116 Istanbul, Turkey Abstract – This paper reports on the assessment of morphological (macro and micro) and anatomical characters of seeds of Romulea taxa distributed in Turkey with the use of one- way analysis of variance, cluster analysis and principal component analysis. Morphologi- cal characteristics such as size, shape, color and surface of seeds were examined with the use of light and scanning electron microscopes. Thicknesses of testa and phytomelan lay- er, sizes of embryo in seeds were studied anatomically. The outcomes revealed that taxa were similar in some aspects such as color and shape of seeds. However, seed size, thick- ness of testa and phytomelan layer, shape of the epidermal cells in testa and sizes of em- bryo were different among taxa, and have taxonomic value in the distinction of these taxa from each other. In addition, the seed surfaces were more or less different for the exam- ined taxa on an interspecifi c level. Consequently, seed morphology and anatomy with a few exceptions demonstrated diversity and they had taxonomic importance in terms of distinguishing among species. Keywords: Anatomy, morphology, phytomelan, Romulea, scanning electron microscopy, seed, Turkey Introduction Romulea Maratti belongs to the subfamily Crocoideae (Ixioideae) in the family Iridace- ae. The Ixioideae is the biggest subfamily of Iridaceae, with more than 800 species of 30 genera (ISIK and DONMEZ 2007). The Ixoideae is a consistent subfamily in terms of morpho- logic and anatomic features (RUDALL and GOLDBLATT 1991, OZDEMIR et al. 2011). The Rom- ulea genus originated from South Africa, and spreads over a wide area, including the Medi- terranean zone, has more than 90 species in a range of distribution areas. The genus Romulea is represented by 7 taxa in Turkey, all of which are listed in table 1 (MARAIS 1984, EROL and KUCUKER 2003). * Corresponding author, e-mail: biology_61@hotmail.com Copyright® 2015 by Acta Botanica Croatica, the Faculty of Science, University of Zagreb. All rights reserved. KARAİSMAİLOĞLU M. C. 32 ACTA BOT. CROAT. 74 (1), 2015 In the classifi cation of Romulea species sexual parts such as anther, fi lament and stigma are often used to separate the taxa from each other. However, classifi cation of Romulea taxa in Turkey as well as in Sicily and South-West Europe is diffi cult because of the frequent emergence of gynodioecism, which is the presence of only hermaphrodite or female fl ow- ers in a population. Namely, if there are only female fl owers on the plants, Romulea taxa look morphologically similar (DAVIS 1984, MORET et al. 1992, MORET et al. 1993). This situ- ation leads to some problems in their classifi cation from time to time, thus additional char- acters that will support the existing identifi cation are needed in the classifi cation of Turkish Romulea taxa. Seeds and small fruits have typical morphological features, such as shape, colour, di- mension, microstructure (including ultrastructure) and they may provide valuable contribu- tions to plant taxonomy. In a practical sense, many signifi cant data gathered from seed or fruit epidermal surface analyses are acquired with scanning electron microscope (SEM). These surface features include micromorphological characteristics, such as shape of the cells, seed surface ornamentation and epicuticular protrusions. These features demonstrate a large diversity from subspecies to family level and they are not readily changed under the infl uence of habitat conditions (BARTHLOTT 1981). However, anatomical features are usually as helpful as morphological features for plant diagnostics, and they often are useful in the separation of closely related taxa (KARAMIAN et al. 2012, AKALIN-URUSAK and KIZILARSLAN 2013). The signifi cance of morphology and anatomy in the classifi cation of taxa in the Iridaceae has been revealed by various studies (MARAIS 1984, EROL and KUCUKER 2003, ISIK and DON- MEZ 2007). Accordingly, the main objective of this study is to investigate morphological and anatomical features of the seeds of the Romulea species native to Turkey with light micro- scope (LM) and SEM in order to discover differences and specifi cities of the seed morphol- ogy and anatomy of examined taxa with the use of statistical analysis. Exhaustive descrip- tion of seed structures of the examined species are offered in this paper for the fi rst time. Materials and methods Plant material Seeds of 7 taxa, which had been gathered from native populations in Turkey and culti- vated at the Alfred Heilbronn Botanical Garden in Istanbul University, were utilized for Tab. 1. Examined taxa and their origin. No Taxa Locality (Origin) R1 R. bulbocodium (L.) Seb. & Mauri var. bulbocodium Izmir, Çeşmealti R2 R. bulbocodium (L.) Seb. & Mauri var. crocea (Boiss. & Heldr.) Baker Antalya, Manavgat R3 R. bulbocodium (L.) Seb. & Mauri var. leichtliniana (Heldr. ex HaI.) Bég. Hatay, Samandag R4 R. tempskyana Freyn Antalya, Falezler R5 R. ramifl ora Ten. subsp. ramifl ora Balikesir, Patria R6 R. linaresii ParI. subsp. graeca Bég. Istanbul, Kurtköy R7 R. columnae Seb. & Mauri subsp. columnae Izmir, Yamanlar MORPHOLOGICAL AND ANATOMICAL STUDIES OF ROMULA TAXA SEEDS ACTA BOT. CROAT. 74 (1), 2015 33 morphological and anatomical observations. Investigations were performed with 10 ripe seeds for each taxon. The taxa examined and their origins are given in table 1. Morphological and anatomical analyses Macromorphological features such as the shape, size, raphe presence or absence and colour of the seeds were examined using an Olympus ZS51 stereomicroscope and Kamer- am Imaging Software. For micromorphological observations of seed surface ornamenta- tion, specimens were prepared for electron microscopy by mounting with silver adhesive on the stub, coated with gold, and analysed with a JEOL Neoscope-5000 scanning electron microscope. For anatomical features such as testa thickness, phytomelan layer thickness, shape of epidermal cells and embryo size, cross sections were taken with a fully automatic micro- tome (Thermo Shonda Met Finesse) from the middle of the seeds. Afterwards, they were passed through a variety of alcohol and xylene series, and stained with haematoxylin (Har- ris-RRSP67-E) in a staining device (ASC 720 Medite), and were covered with Entellan for anatomical structures to be examined (INCEER et al. 2010). Anatomical characters were ob- served using an Olympus CX21FS1 microscope and Kameram Imaging Software. The terminology of morphological and anatomical characters was carried out in accord- ance with STEARN (1985). Statistical analyses Data of studied parameters were performed with the SPSS computer program. The Duncan multiple-range test was utilized to determine the statistical signifi cance of differ- ences among the values obtained for different taxa. Grouping of taxa was carried out utiliz- ing the clustering analysis method (UPGMA) in accordance with quantitative characters in Tab. 2. In addition, principal component analysis (PCA) ordination and similarity matrix were performed based on some morphological and anatomical characters of seeds such as seed dimensions, testa thickness, phytomelan layer thickness and embryo sizes. Results Seed size, raphe and shape characters of taxa were evaluated macromorphologically and differences among the studied taxa were observed (Tab. 2). In seed dimensions, R6 and R7 had the greatest differences among the examined species. Their values range from 2.04 mm to 4.54 mm for length and from 2.88 mm to 3.67 mm for width, respectively. Colour of seeds in all cases is orange, but colour tones are different in R1 (darker). However, colour of seeds is determined to be unimportant in the distinction of the taxa of Romulea exam- ined. The shape of seeds in these taxa indicates major differences. Likewise, the ratio val- ues of the seed sizes show variations, which vary between 0.60 and 1.28. Seeds are ovate in R1 (0.60), R2 (1.28), R4 (0.72), R5 (1.10), R6 (0.70), and elliptical in R3 (1.25), R7 (1.23). However, the raphes of the taxa also differ. The raphe is very prominent in R4 and R7, com- pared to other species (Fig. 1). Seed surfaces of the studied taxa exhibited diversity of surface ornamentation (Fig. 2 and Tab. 2). The seed surface ornamentation, which is a useful character in the separation of KARAİSMAİLOĞLU M. C. 34 ACTA BOT. CROAT. 74 (1), 2015 Ta b. 2 . M or ph ol og ic al a nd a na to m ic al fe at ur es in s ee ds o f t he T ur ki sh R om ul ea ta xa . R es ul ts re pr es en t m ea n va lu es ± s ta nd ar d de vi at io n; m ea ns w ith d if - fe re nt le tte rs a re s ig ni fi c an t a t p = 0 .0 5 le ve l ( D un ca n’ s m ul tip le -r an ge te st ), fo r t ax a ab br ev ia tio ns s ee T ab . 1 ; L – le ng th , W – w id th . Ta xa C ol ou r Sh ap e Se ed s iz e R at io (L /W ) Se ed su rf ac e or na m en - ta tio n Te st a th ic kn es s (μ m ) Ph yt om el an la ye r th ic kn es s (μ m ) Te st a ep id er m al ce lls E m br yo s iz es L (m m ) W (m m ) L (μ m ) W (μ m ) R 1 D ar k or an ge O va te - Tr an sv er se 2. 14 ± 0. 15 de 3. 53 ± 0. 11 c 0. 60 R et ic ul at e 21 8. 84 ± 10 .7 0a b 54 .2 0± 5. 81 d U nd ul at ed 29 8. 38 ± 11 .3 2d 22 4. 47 ± 15 .3 5c d R 2 O ra ng e O va te - L at e 3. 87 ± 0. 12 a 3. 02 ± 0. 21 de 1. 28 R et ic ul at e- Fo ve at e 21 3. 05 ± 8. 64 b 86 .0 1± 6. 37 b C ru sh ed 31 7. 91 ± 7. 55 c 28 9. 25 ± 11 .6 5b R 3 O ra ng e E lli pt ic al - L at e 3. 04 ± 0. 28 c 2. 43 ± 0. 08 e 1. 25 R et ic ul at e- A re ol at e 81 .9 3± 10 .5 6e 54 .1 1± 10 .6 9c d C ru sh ed 17 5. 98 ± 35 .9 9f 15 6. 82 ± 21 .2 4e R 4 O ra ng e O va te - L at is si m e 2. 51 ± 0. 17 d 3. 48 ± 0. 04 b 0. 72 R et ic ul at e 23 8. 31 ± 5. 58 a 77 .0 3± 11 .3 8b c C ru sh ed 24 6. 04 ± 34 .0 8e 19 6. 41 ± 18 .6 3d R 5 O ra ng e O va te - L at e 3. 56 ± 0. 08 b 3. 21 ± 0. 12 d 1. 10 R et ic ul at e- A re ol at e 18 3. 21 ± 14 .6 3c 56 .0 1± 9. 41 cd C ru sh ed 38 8. 80 ± 11 .7 9a 32 8. 60 ± 12 .4 2a R 6 O ra ng e O va te - D ep re ss e 2. 04 ± 0. 16 e 2. 88 ± 0. 19 de 0. 70 R et ic ul at e 13 5. 79 ± 19 .2 5d 99 .3 2± 3. 54 a C ru sh ed 35 1. 44 ± 17 .4 3b 28 1. 12 ± 33 .4 9b c R 7 O ra ng e E lli pt ic al 4. 54 ± 0. 49 a 3. 67 ± 0. 38 a 1. 23 A lv eo la te 15 6. 53 ± 20 .9 1c d 63 .6 3± 2. 93 c Fl at 29 6. 63 ± 12 .6 8d 24 5. 33 ± 10 .6 6c MORPHOLOGICAL AND ANATOMICAL STUDIES OF ROMULA TAXA SEEDS ACTA BOT. CROAT. 74 (1), 2015 35 taxa, varies from slightly (R3) or intensive- ly (R2) reticulate to alveolate (R7). How- ever, seeds of R5 have a vaguely reticulate- areolate sculpture, while there are elevated protrusions on the seed surface of R1 which is distinguished by these protrusions from R4 and R6 (Fig. 2). On the other hand, the seed surface patterns of R4 and R6 are not signifi cant in terms of the sepa- ration of taxa (Tab. 2). The anatomical characteristics of the seeds of Turkish Romulea are given in ta- ble 2. Accordingly, the mean values of tes- ta thickness vary from 81.93 μm to 238.31 μm, and this character varied signifi cantly among the examined taxa. A thick testa is noted in R4, whereas the testa in R3 is thin (Tab. 2). In other respects, shapes of epi- dermal cells of testa differ in the studied taxa (Tab. 2), which have elevated-fl at, crushed and undulating cells in unequal or equal forms, and thick or thin walls in cross sections (Fig. 3). In addition to these fi ndings, embryo sizes of the seeds have been shown to range from 175.98–388.80 μm to 156.82– 328.60 μm, and embryos in seeds formed oval (R4 and R5), globular (R2 and R3) and pro- longed (R1, R6 and R7) in shapes (Fig. 3). Fig. 1. Turkish Romulea seeds; a: R1, b: R2, c: R3, d: R4, e: R5, f: R6, g: R7; for taxa abbreviations see Tab. 1; scale bars = 1 mm. Fig. 2. Seed surfaces of Romulea taxa; a: R1; Reticulate, b: R2; Reticulate-Foveate, c: R3; Reti- culate -Areolate, d: R5; slightly Reticulate-Areolate, e: R7; Alveolate; for taxa abbreviations see Tab. 1. The arrow shows protrusion on the seed surface. KARAİSMAİLOĞLU M. C. 36 ACTA BOT. CROAT. 74 (1), 2015 An unweighted pair group method with arithmetic mean (UPGMA) dissimilarity clus- tering dendrogram for Turkish Romulea taxa according to the examined quantitative char- acters in Tab. 2 is given in Fig. 4. Accordingly, branches in the dendrogram are divided into two main clads and sub-clads. R7 composes the fi rst main branch; other taxa make up the second main branch. Also, R7 differs clearly from other taxa in terms of characters deter- Fig. 3. Cross section structures of Turkish Romulea seeds; 1: overall appearance; anticlinal cell types: 2: fl at cell (R7), 3: crushed cell (R2, R3, R4, R5 and R6), 4: undulated cell (R1), types of embryo, 5: oval (R4 and R5), 6: prolonged (R1, R6 and R7), 7: globular (R2 and R3); for taxa abbreviations see Tab. 1; e – embryo, en – endosperm, sr – storage reserve, t – testa, ec – epidermal cells of testa, fc – fl at cell, ph – phytomelan layer, cc – crushed cell, uc – undu- lated cell; white scale bars = 100 μm, black scale bars = 50 μm. Fig. 4. UPGMA clustering of Romulea taxa in Turkey based on morphological and anatomical characters of the seeds; for taxa abbreviations see Tab. 1. MORPHOLOGICAL AND ANATOMICAL STUDIES OF ROMULA TAXA SEEDS ACTA BOT. CROAT. 74 (1), 2015 37 mined in Tab. 2. Furthermore, PCA ordination and similarity matrix based on some mor- phological and anatomical characters of seeds such as seed dimensions, testa thickness, phytomelan layer thickness and embryo sizes are presented in Fig. 5 and Tab. 3. Among the studied taxa, the closest and the most distant species are determined. According to these results, R6 and R2 are the most closely related species (percentage similarity: 0.789), R6 and R7 are the most distantly related species (percentage similarity: –0.169) (Tab. 3 and Fig. 5). In addition, the cumulative variance value of principal components achieved 63.14 % (axis 1: 32.29%, axis 2: 30.85%). Fig. 5. Principal component analysis of Romulea taxa in Turkey based on morphological and anatomical characters of the seeds; for taxa abbreviations see Tab. 1. Tab. 3. Similarity matrix among the Turkish Romulea taxa based on some morphological and ana- tomical characters of seeds; for taxa abbreviations see Tab. 1. Taxa R1 R2 R3 R4 R5 R6 R7 R1 1 – – – – – – R2 0.310 1 – – – – – R3 0.070 0.514 1 – – – – R4 –0.0440 0.383 0.633 1 – – – R5 0.070 0.514 0.514 0.383 1 – – R6 0.292 0.789 0.310 0.450 0.310 1 – R7 0.070 0.029 0.271 –0.1180 0.029 –0.1690 1 KARAİSMAİLOĞLU M. C. 38 ACTA BOT. CROAT. 74 (1), 2015 Discussion The morphological (macro and micro) and anatomical characters of seeds frequently indicate major differences among taxa of the same genus or family, and these differences provide a signifi cant contribution to delimiting various taxonomic levels of taxa and to the establishment of taxonomic relationships. Although reliability and sustainability of seed properties in classifi cation are known (DAHLGREN and CLIFFORD 1982, GRILLI CAIOLA et al. 2010), no researches appeared to have been performed so far concerning seed morphology and anatomy of Romulea taxa in Turkey. In this investigation, macromorphological features such as size, shape and raphe char- acters of seeds of Romulea taxa in Turkey have indicated the presence of variations among the examined taxa. Although it is reported as being only globular in the Flora of Turkey and Aegean Islands by MARAIS (1984), seed shape in almost all of the examined Romulea taxa varied (excluding seed shapes of R2 and R5) (Tab. 2). The seed sizes of Romulea taxa are quite variable. The broadest seeds are found in R7, and the smallest seeds in R6. Both of these measurements are considerably different from those obtained from other examined taxa. These variations in the seed shape are compatible with the diagnostic characters in Flora of Turkey (DAVIS 1984) for the Romulea species. In contrast to the fi ndings of MARAIS (1984), the seed colour of Turkish Romulea taxa has been found to be orange and its tones rather than brown, and the colour of seed has no use taxonomically. One of the remarkable characters is also the presence of raphes on seeds, and their size. The raphes of R4 and R7 are more pronounced than on other examined taxa, and there is no prominent raphe in R1 (Fig. 1). At the same time, the sizes of seeds having a raphe are generally smaller than those do not have a marked raphe. This result is attributable to the principle that wind-dispersed seeds should be in small size and should bear raphe-like structures (TEBBITT 2005). The seed morphology has not been suffi ciently utilized in the taxonomy of Iridaceae family (EROL et al. 2006). Results obtained from this research indicate that some macro- morphological features, particularly the shapes and sizes of seeds of Romulea taxa reveal as distinctive characters in terms of classifi cation, because of their diversity among taxa, so they may be used as supporting characters in taxonomic studies. Seed surface structures have been utilized for some purposes such as the solution of systematic problems, interpretation of evolutionary interactions, and illumination of the adaptive features of the seed surface (HEYWOOD 1971, SULAIMAN 1995). In this investiga- tion, the surface patterns range from slightly (R3) or intensively (R2) reticulate to alveolate (R7) in the examined taxa. R5 has very weak reticulate-areolate surface and the seed sur- face of R1 has visible raised protrusions (Fig. 2). And so, the surface patterns of seed coats are found helpful in taxonomy for some Romulea. Nonetheless, variations in the obtained results indicate that the seed surface characters support the frequently used characters in the systematics of the genus (according to DAVIS 1984) (except for R4 and R6), and can con- tribute to classifi cation of Turkish Romulea species. The testa thicknesses and epidermal cell types of the seeds can provide precious data about phylogenetic classifi cation of fl owering plants (CORNER 1976). In this investigation, the differences in anatomical features such as testa thickness, phytomelan thickness, shape of epidermal cells of testa and embryo size of the seeds of Turkish Romulea have been identifi ed. The most distant species are R3 (81.93 μm) and R4 (238.31 μm), while the other species examined are located between 81.93 μm and 238.31 μm. Likewise, the epidermal MORPHOLOGICAL AND ANATOMICAL STUDIES OF ROMULA TAXA SEEDS ACTA BOT. CROAT. 74 (1), 2015 39 cells of testa indicate difference in Turkish Romulea species. They form different shapes such as undulating (R1), polygonal crushed (R2–R6) and fl at (R7) in the cross sections of the seeds of the examined taxa (Tab. 2). Accordingly, epidermal cells can be used in the separation of R1 and R7 from other taxa, and can be of some use in the taxonomy of exam- ined taxa. In this regard, the results obtained are consistent with a previous similar study in Iridaceae (GRILLI CAIOLA et al. 2010). The taxonomic signifi cance of phytomelan-covered seeds was explained by HUBER (1969), and the presence-absence and thickness (if any) of phytomelan layer are of notable taxonomic signifi cance in the distinction of taxa in the Iridaceae family (DAHLGREN and CLIFFORD 1982). The thickness of the phytomelan layer in Turkish Romulea species ranged from 54.11 μm to 99.32 μm and changed signifi cantly in examined Romulea species (Tab. 2). This fi nding attributed as the thickness of phytomelan layer can be useful in the classifi - cation of Turkish Romulea species because of differentiated thickness in almost all of the examined taxa in this investigation (Tab. 2). The embryos of the Turkish Romulea taxa exhibit a wide variety in terms of shapes and sizes (Tab. 2). While the embryo in R2 and R3 is usually swollen and well arranged, the embryo in other examined taxa is more or less elongated (as if not well developed).The an- atomical fi ndings of this investigation are compatible with those of GRILLI CAILO et al. (2010), who have made a study with similar variables including anatomical features on some Crocus seeds. One of the most important characters which distinguish R1, R4 and R6 from each other is the colour and length of fi laments (DAVIS 1984). However, it is impossible to separate the taxa from each other in the case of gynodioecism in one of the populations of these taxa since there will be only female fl owers. In this investigation, seed shape, size and presence of raphe on seeds morphologically; thickness of phytomelan layer, embryo size anatomi- cally are applicable characters in distinguishing these taxa. UPGMA of cluster analysis was used to evaluate the morphological and anatomical features of the seeds among the analyzed taxa. In this dendrogram, branches that are mark- edly dissimilar to other taxa were created for R1 and R7. This fi nding can be attributed to R1 and R7 being different from the other analyzed taxa in terms of the studied characters. Generally, the branches contained nearly related taxa are compatible with the traditional taxonomic rank of Turkey’s Romulea species. In other words, the morphological and ana- tomical features of the seeds supported the characters used in the distribution of Romulea species in the fl ora of Turkey (according to DAVIS 1984). Principal component analysis can be helpful in giving data about the variability of quan- titative characters. The obtained cumulative variance values of principal components show that the studied characters in Turkish Romulea taxa can be of use in explaining the differ- ences among the taxa because of high variance value. In addition, the morphological and anatomical features of the seeds chosen for PCA to evaluate the characteristics that are sig- nifi cant in description change among the analyzed specimens (Tab. 3). Likelihood ratios among the taxa were determined. The closest relationship was seen between R6 and R2, additionally the most distant relationship was found between R6 and R7. These results indi- cate that a similarity matrix can give information about the classifi cation of Turkish Romu- lea taxa if there is gynodioecism in a population. KARAİSMAİLOĞLU M. C. 40 ACTA BOT. CROAT. 74 (1), 2015 In conclusion, this investigation supports the use of seed surface, shape, size and the cross section of seed such as testa and phytomelan layer thickness, shape of testa epidermal cells and embryo sizes as the distinctive characters in the classifi cation within the Romulea taxa in Turkey. Acknowledgements The author thanks Dr. Osman Erol for supplying seeds of the studied taxa and Veysel Süzerer and Almıla Çiftçi for technical assistance. References AKALIN-URUSAK, E., KIZILARSLAN, C., 2013: Fruit anatomy of some Ferulago (Apiaceae) species in Turkey. Turkish Journal of Botany 37, 434–445. BARTHLOTT, W., 1981: Epidermal and seed surface characters of plants: Systematic applica- bility and some evolutionary aspects. Nordic Journal of Botany 1, 345–355. CORNER, E. J., 1976: The seeds of the dicotyledons. Vol. 2. Cambridge University Press, Cambridge. DAHLGREN, R. F., CLIFFORD, H. T., 1982: The monocotyledons: A comparative study, Aca- demic press, London. DAVIS, P. H., 1984: Flora of Turkey and the East Aegean Islands, Vol. 8. Edinburgh Univer- sity Press, Edinburgh. EROL, O., KUCUKER, O., 2003: Morpho-anatomical observations on three Romulea (Iridace- ae) taxa of Turkey. Bocconea 16, 607–613. EROL, O., UZEN, E., KUCUKER, O., 2006: Preliminary SEM observations on the seed testa structure of Gladiolus L. species from Turkey. International Journal of Botany 2, 125– 127. GRILLI CAIOLA, M., LEONARDI, D., CANINI, A., 2010: Seed structure in Crocus sativus L. X, C. cartwrightianus Herb., C. thomasii Ten., and C. hadriaticus Herb. at SEM. Plant Systematics and Evolution 285, 111–120. HEYWOOD, W. H., 1971: Scanning elentron microscopy. Academic Press, London. HUBER, H., 1969: Die samenmerkmale und Verwandtschafts-verhaltnisse der Liliifl orae. Mitteilungen der Botanischen Staatssammlung München 8, 219–538. INCEER, H., BAL, M., CETER, T., PINAR, N. M., 2010: Fruit structure of 12 Turkish endemic Tripleurospermum Sch. Bip. (Asteraceae) taxa and its taxonomic implications. Plant Systematics and Evolution 298, 845–855 ISIK, S., DONMEZ, E. O., 2007: Pollen morphology of the Turkish Romulea Maratti (Iridace- ae). Turkish Journal of Botany 31, 171–182. KARAMIAN, R., BEHJOU, A. M., RANJBAR, M., 2012: Anatomical fi ndings of Onobrychis sect. Heliobrychis (Fabaceae) in Iran and their taxonomic implications. Turkish Journal of Botany 36, 27–37. MARAIS, W., 1984: Romulea Maratti In: Davis PH (ed.), Flora of Turkey and the East Ae- gean Islands, vol. 8, pp. 438–441. Edinburgh University Press, Edinburgh. MORPHOLOGICAL AND ANATOMICAL STUDIES OF ROMULA TAXA SEEDS ACTA BOT. CROAT. 74 (1), 2015 41 MORET, J., BARI, A., LE THOMAS, A., GOLDBLATT, P., 1992: Gynodioecy, herkogamy and sex- ratio in Romulea bulbocodium var. dioica (Iridaceae). Evolutionary Trends in Plants 6, 99–109. MORET, J., BARI, A., LE THOMAS, A., 1993: Evolution of herkogamy and gynodioecy in Mo- roccan species of Romulea (Iridaceae). Plant Systematics and Evolution 184, 241–257. OZDEMIR, C., BOZDAG, B., AKYOL, Y., SEN, U., SEPET, H., YETISEN, K., 2011: Morphological and anatomical investigations of Romulea bulbocodium var. bulbocodium and Romulea bulbocodium var. leichtliniana (Iridaceae). Thaiszia Journal of Botany 21, 65–72. RUDALL, F. L. S., GOLDBLATT, P., 1991: Leaf anatomy and phylogeny of Ixioideae (Iridace- ae). Botanical Journal of the Linnean Society 106, 329–345. SULAIMAN, I. M., 1995: Scanning electron microscopic studies seed coat patterns of fi ve en- dangered Himalayan species of Meconopsis (Papaveraceae). Annals of Botany 76, 323– 376. STEARN, W. T., 1985: Botanical Latin: history, grammar syntax, terminology and vocabu- lary. David & Charles, London. TEBBITT, M. C., 2005: Begonias, cultivation, identifi cation and natural history. Timber Press, Portland.