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(South Poland) Endemic Species Erysimum Pieninicum (Zapa£.) Paw£

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(South Poland) Endemic Species Erysimum Pieninicum (Zapa£.) Paw£ Vol. 76, No. 2: 127-132, 2007 ACTA SOCIETATIS BOTANICORUM POLONIAE 127 SEM OBSERVATIONS OF POLLEN GRAINS, FRUITS AND SEEDS OF THE PIENINY MOUNTAINS (SOUTH POLAND) ENDEMIC SPECIES ERYSIMUM PIENINICUM (ZAPA£.) PAW£. (BRASSICACEAE) IRMINA MACIEJEWSKA-RUTKOWSKA1, LESZEK BEDNORZ2, TOSHIYUKI FUJIKI3 1 Department of Forest Botany, August Cieszkowski Agricultural University Wojska Polskiego 71 D, 60-625 Poznañ, Poland e-mail: [email protected] 2 Department of Botany, August Cieszkowski Agricultural University Wojska Polskiego 71 C, 60-625 Poznañ, Poland 3 Nagoya University, Graduate School of Environmental Studies, School of Sciences Furo-cho, Chikusa-ku Nagoya City, 464-8601 Japan (Received: June 22, 2006. Accepted: August 31, 2006) ABSTRACT Erysimum pieninicum (Zapa³.) Paw³. (Brassicaceae) is an endemic species, growing only in Pieniny Mts. The aim of the presented work was to investigate its pollen morphology as well as its ultrastructural traits of pericarp and seed-coat. Pollen grains of this species were of small size, 3-zonocolpate with reticulate ornamentation. The external siliqua surface was of rugose sculpture, covered by numerous, stellate hairs. The internal surface of the fruit was naked and characterized by striate sculpture. The seed-coat ornamentation of E. pieninicum was of bli- ster type. KEY WORDS: Erysimum pieninicum, endemic species, pollen morphology, fruit morphology, seed morphology, SEM. INTRODUCTION all Polish Erysimum species with the use of light microsco- pe, was not able to clearly separate some taxa. The use of According to Koch and others (2003) Erysimum is taxo- electron microscope (SEM) may resolve many systematic nomically one of the most difficult genus of the mustard questions. For example the study of Koul and others (l.c.) family (Brassicaceae) and extensive phylogenetic studies on the Brassica and allied genera supported the use of seed on this taxon are necessary. It comprises many taxa of coat microsculpture pattern as a parameter for species uncertain status and the total number of described species identification. Detailed investigations on the anatomy of oscillates between 80 and 180 (Latowski 1975; Koch et al. the chazal endosperm cyst of 11 genera of Brassicaceae l.c). Whereas the taxonomy of the genus has traditionally (Brown et al. l.c.) also showed the potential taxonomic va- been based on a rather small number of characters, such as lue of this feature. annual versus perennial life cycles, some macroscopic tra- The ultrastructural patterns of the pericarp in mustard fa- its of flowers and fruits, leaf architecture or trichome mor- mily have not been recognized yet. Similarly, there still phology, their application does not always allow to distin- lack SEM studies of pollen morphology of most the Bras- guish some closely related species (Latowski 1985; Ball sicaceae species. According to the authors knowledge, 1993). only pollen grains of several species of Erysimum were ob- With reference to different representatives of Brassicace- served under the scanning electron microscope (see An- ae, the search for distinct and unequivocal indicators chev and Deneva 1997; Perveen et al. 2004; Halbritter prompted some botanists to examine seed morphology and 2006). anatomy (e.g. Vaughan and Whitehouse 1971; Latowski Hence, the aim of the present work was to investigate the 1975; Garnock-Jones 1991; Koul et al. 2000; Brown et al. pollen morphology of Erysimum pieninicum, as well as the 2004; Zeng at al. 2004). As regards seed anatomy, Vau- ultrastructural traits of siliqua pericarp and seed-coat. ghan and Whitehouse (l.c.) noticed that testa may be of Since the taxonomic revision carried out by Paw³owski greatest value in determining taxonomic relationships wi- (1946), this taxon has been distinguished in the rank of thin the mustard family. However, Latowski (l.c.) analy- species, however sometimes it is included in Erysimum zing both morphological and anatomical traits of seeds of hungaricum sensu lato, together with E. wahlenbergii (Ball 128 POLLEN GRAINS, FRUITS AND SEEDS OF ERYSIMUM PIENINICUM Maciejewska-Rutkowska I. et al. 1993). Among the nine species of wallflower noted in Po- Siliqua morphology (Fig. 3) land, this one belongs to the rarest. It is an endemic spe- The fruit morphology of Erysimum pieninicum was de- cies, growing only in Pieniny Mts., altitude 500-790 scribed with the use of LM by Latowski (1975). According m a.s.l., on four sites. The species is listed in Bern Conven- to this author the siliquas of this species are on the average tion on the Conservation of European Widelife and Natural 40-60 mm long and 1.5 mm wide, linear, with rhombic ou- Habitats and it is protected by law in Poland. Still in 2001 tline of cross-section. The base of fruit is rounded and the Kamierczakowa and Zarzycki considered E. pieninicum to apex is acute, with the beak, consisting stigma, 1.1-2.0 mm be a threatened species of the Polish vascular flora, while long. The stigma is distinctly bilobate, wider than beak. Zarzycki and Szel¹g (2006) include it to the group of rare Fruit usually sparsely hairy, sometimes densly. Hairs stel- plants in Poland. late. Valves with only midvein distinct. The septum is membranous, naked, of the same thickness, as the valves. The current study of the fruit ultrastrastructure revealed MATERIALS AND METHODS the external surface of the valve to be rugose with irregu- lar, relatively shallow wrinkles. The cells of the outer layer In 2005 the flowers and the fruits of Erysimum pienini- were izodiametric, ± polygonal or rounded, with the sligh- cum were collected from two natural localities of Pieniny tly protruding latteral walls and with somewhat concave Mts: the hill of the Czorsztyn castle (E 20°19, N 49°26, outer ones, but the cell outlines were rather weakly mar- 550 m a.s.l.) and slopes of Flaki Mt. (E 20°21, N 49°25, ked. Instead, at high magnification (at least 200×) the layer 790 m a.s.l.). Two samples of pollen grains (ripe anthers of striate cuticule, consisting of the striae running parallel were taken only) were acetolysed according to Erdt- to the fruit axis, was visible. The surface was covered by mans method (1952), with some insignificant modifica- stellate, three- or four-armed, rarely two-armed hairs; with tions (Tomlik-Wyremblewska 1995). The samples of fruits the diameter about 100-200 µm. The arms were curved and and seeds (properly developed and ripe) were not prepared, gradually tapered, with acute ends. The hairs of pimple te- but only cleaned. The use of scanning electron microsco- xture, with granules of different size. Additionally, there pes (JEOL JSM 6300 and LEO 435 VP) allowed to investi- were scattered rod-like elements, no more than 10 µm high gate the details of exine sculpture and the silique and seed and stomata, ±15 µm long. The inner surface of valve was ultrastructure. Light microscopy (BIOLAR 2308) was ma- of striatae sculpture. The cells were izodiametric, ± roun- inly carried out to study the exine structure and quantitati- ded, with outer walls convex and lateral ones concave and ve characters of pollen grains. In all 60 pollen grains of often undulate-sinuate. The inner surface was also covered two samples were observed. The pollen terminology fol- by thin layer of rugulate cuticule, without hairs or pores. lows Hoen (1999) and Halbritter and others (2006). With The septum surface was rugose. The outline of cells of the regard to the seeds and fruits, terminology of Barthlott outer layer was rather poorly marked. There was no pube- (1981), based on SEM observations of epidermal and seed scence on the surface of septum, with the exception of its coat surface in 5000 species of seed plants, was applied. lateral edge, sometimes with the single, stellate hairs. Seed morphology (Fig. 4) RESULTS According to Latowski (1975), the seeds of the species Pollen morphology (Figs 1-2) of Erysimum develop from campylotropous ovules. The mature seeds of E. pieninicum are ±1,81-2,40 mm long and In this study pollen grains of Erysimum pieninicum were 0,81-0,90 mm wide; laterally flattened, variable in outline: of small size; on average length of polar axis (P) was 18.00 oblong elliptical, oval or almost rectangular, with gently µm (with the whole range 15.00-21.00 µm) and equatorial rounded angles. The seed apex is straight or obliquely trun- axis (E) 17.25 µm (with the whole range 13.50-19.50 µm). cate, with the protruding radicle. At the same level there is P/E value oscillated from 0.91 to 1.30, with the mean value also visible a blackish spot, enclosing hilum in the form of 1.05. The grains were most often spheroidal in shape fine membrane, sometimes with the rest of funicle. The ba- (54.6% of observed grains), sometimes prolate spheroidal se of seed may be obliquely acute or gently rounded, with (33.3%), rarely subprolate (7.6%) or oblate spheroidal the patch-like wing, 300-500 µm long. Seed-coat 50-100 (4.5%); with equatorial outline circular or lobate. µm thick, consisting of 3-4 layers. Testa cells are polygo- Three-zonocolpate, sporadically four-zonocolpate pollen nal in outline, at cross-section with mucilaginous excre- grains were observed. Colpi were usually comparatively scences, mushroom-shaped, up to 20 (-35) µm high. narrow and deeply sunken in exine, with acute ends. Aper- The present investigation of seed ultrastructure of Erysi- ture length was more than 4/5 length of polar axis. Colpus mum piennicum revealed the blister type of seed-coat margin was regular, membrana colpi granulate. sculpture. The cells of the testa were izodiametric, penta- Exine always consisted of two layers, well visible in LM. or hexagonal, sometimes almost circular in outline.
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