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OCCURRENCE OF ONEROW YELLOWCRESS MICROPHYLLUM BOENN. RCHB. IN THE ILANKA RIVER LUBUSZ LAND

A C, M M, I J

A. Czarna, M. Morozowska, Department of Botany, Poznań University of Life Sciences, Wojska Polskiego C, - Poznań, Poland, e-mail: [email protected], [email protected] I. Jędrzejczyk, Departm ent of Genetics, Physiology and Biotechnology, Laboratory of Molecular Biology and Cytometry, University of Technology and Life Sciences, Kaliskiego Ave. , - Bydgoszcz, Poland, e-mail: [email protected]

(Received: March , . Accepted: April , )

ABSTR ACT. The paper presents three new localities of Nasturtium microphyllum reported on the Ilanka River near Rzepin. In one of the localities onerow yellowcress specimens were in the fl owering and fruiting stages, while in the two other localities were fl owering, but they were not bearing fruits. Fruiting specimens were identifi ed on the basis of seed sculpture traits, while fl owering plants – by fl ow cytometry, based on the genome size.

KEY WORDS: Nasturtium microphyllum, protected plants, the Ilanka, distribution, SEM, genome size

INTRODUCTION IDENTIFICATION METHODS

In Poland onerow yellowcress Nasturtium microphyl- Onerow yellowcress may be most reliably identifi ed lum (Boenn.) Rchb. () is covered by on the basis of the arrangement of seeds distributed in strict species protection (R M... a single row on both sides of the fruit septum, due to ) and it has been included in the Polish Red Book which fruits are narrow. In watercress seeds are found of Plants, where it is classifi ed as a vulnerable taxon, in two rows and fruits are wider, while the third, hybrid VU (S ). To date a total of ten localities species forms rudimental siliques, typically with a limited have been reported in Poland, while at present only number of sterile seeds (C et . ). A valuable seven localities of this species exist, i.e. Mydlniki near diagnostic tool is also provided by the reticullate seed Kraków (T ), Makrosice near Gubin (C sculpture, visible under a scanning electron microscope and M ), two localities near Kraków (SEM). On seeds of onerow yellowcress the sculpture is (K and S ), Bielinek, the Odra river fi nely reticulate with over cells on one side of the valley near Chlewice (S ), Zieleniewo near seed (Figs A, B), on seeds of watercress seed surface Kołobrzeg and Babimost near Zielona Góra (C and cells are large, ranging in number from to , while M ). The distribution of N. microphyl- on seeds of the hybrid the number of seed sculpture lum in Poland is still relatively little known due to the cells ranges from to (B et . , , recent introduction of this species into Polish keys for N , H and M ). the identifi cation of species (T , R Identifi cation of species from the genus Nasturtium ) and the great morphological similarity to com- in the vegetative state or during fl owering is very dif- mon watercress Nasturtium offi cinale R. Br. , as well fi cult. It may prove helpful to compare leaf outline, as brown watercress Nasturtium ×sterile (Airy Shaw) which in all examined cress species is pinnate. Their Oefelein . morphological variation is manifested fi rst of all in the Onerow yellowcress has practical and medicinal uses shape of individual leafl ets, although it does not ensure similar to those of watercress (http://www.pfaf.org). The an unambiguous taxonomic diagnosis (Fig. ). Flow species mentioned above as well as their hybrid are also cytometry, which was originally used only in medical occasionally cultivated. In the th century watercress research, is a reliable method that provides a correct was commercially grown in . At present leaves of identifi cation of species in the Nasturium genus when these species are used as an ingredient of salads, while the fruits and seeds are absent. Nowadays this method seeds as a substitute of mustard seeds are used to season is applied in plant science by botanists, genetics, bio- meat (P i P ). technologists, plant breeders and seed producers. Flow cytometry has been used mostly to determine genome size in various plant species, to analyse ploidy level, cell cycle and endoreduplication in diff erent plant organs. A. Czarna, M. Morozowska, I. Jędrzejczyk

F. . Carpological diagnostic features of Nasturtium microphyllum (SEM): A, B – seed sculpture; C, D – ultrastructure of internal surface of fruit valve; E, F – microornamentation pattern of fruit septum

The measurement is based on the linear relationship in a nuclear isolation buff er supplemented with a fl uo- between fl uorescence of stained nuclei and DNA con- rescent dye. Total nuclear DNA content in the sample tent. Estimation of plant nuclear DNA content is usually is calculated by the comparison of the sample G peak performed using young and fresh leaves (C et . positions and the internal standard on the histogram. , H et . ). However, plant material Plant or animal cells with accurately estimated DNA such as roots, hypocotyls, fl owers, pollen grains or seed contents should be used as a standard. The amount of can be used for FCM as well (S et . , Ś- C DNA is expressed in picograms (pg) or the number et . ). Flow cytometry analysis is performed of base pairs (bp; pg DNA = . × bp; D on a nuclear suspension, after chopping plant material et . ). Occurrence of onerow yellowcress Nasturtium microphyllum (Boenn.) Rchb. in the Ilanka River (Lubusz Land)

NEW LOCALITIES

An inspiration for the undertaking of fi eld stud- ies along the entire course of the Ilanka River (approx. km) was provided by the discovery, in August , of a new locality of N. microphyllum on section of the river in Rzepin (A). In the course of further studies con- ducted in the same vegetation season at a distance of approx. km from locality A two further localities of the investigated species (B and C) were found, at a distance of approx. m from each other (Fig. ). Watercress specimens growing there were in the fl owering stage and for this reason cytometric analyses were performed to provide accurate identifi cation of species. Material for measurements of nuclear DNA content comprised upper, fully developed leaves of N. microphyllum from individual localities. At each locality they were collected from plants growing at a distance of at least m from one another. Plant material for cytometric analy- ses was prepared following the procedure developed for species from the genus of Nasturtium (M et . ). Herbarium data sheets, one from each locality, and a fruit with seeds collected at locality A were deposited at the herbarium of the Department of Botany, the Poznań University of Life Sciences (POZNB).

A – locality ‘Rzepin ’ In the course of fi eld studies onerow yellowcress was found in the Ilanka River in the section in the area of Rzepin, the Rzepin commune, the Słubice county, the F. . The outline and morphological diff erentiation of Lubuskie province. The occurrence of the investigated Nasturtium microphyllum leaves from sites A, B and C lo- species was limited to the river segment from the dam calized in the Ilanka River at the road running in the north-westerly direction up to

F. . Distribution of Nasturtium microphyllum occurrence along the Ilanka River: – localities with the presence of Nasturtium microphyllum: A – ‘Rzepin ’, B – ‘Rzepin ’, C – ‘Starościn’; – localities without the presence of Nastur- tium microphyllum; – railway track A. Czarna, M. Morozowska, I. Jędrzejczyk the bridge on the river (Fig. ). Onerow yellowcress was Plants growing along the river bed were found only in growing here on both sides of the river bed of approx. layer C (total cover %) and included Nasturtium mi- m in width, fi lled with clean, slow-fl owing and relatively crophyllum ., Phragmites australis ., Ceratophyllum shallow water. The rushes zone on both sides of the river demersum ., Phalaris arundinacea ., Acorus calamus was well-developed. Onerow yellowcress in this locality ., Carex acutiformis ., Lemna minor ., Myosostis was in the fl owering and fruiting stages. Based on SEM palustris ., Potamogeton natans ., Sagittaria sagit- analyses it was found that the reticulate cell pattern on tifolia ., Sparganium ramosum ., Glycera maxima +, the inner surface of the fruit valve provides good di- Sium latifolium +, Solanum dulcamara +, Nuphar lu- agnostic traits for the identifi cation of onerow yellow- teum r, Equisetum limosum r, Filipendula ulmaria r, Iris cress. Cells are narrow, strongly elongated with acute pseudacorus r, Rumex hydrolapatum r. The discussed or rounded endings and with a well-visible secondary Nasturtium species is growing in locality B also in the sculpture in the form of transverse striations, on aver- Nasturtietum microphylli community. age - per cell, and accompanying small wrinkles (Figs C, D). Microstructure of the fruit’s septum surface, C – locality ‘Starościn’ which is also of reticulate pattern, is characterised by Specimens of onerow yellowcress in the locality in strongly elongated cells with slightly rounded, straight Starościn were found on both sides of the Ilanka River or acute endings. Anticlinal walls of cells are straight bed of approx. m in width, in the m long seg- or frequently sinuate. The described characteristics of ment, in the form of distinct patches fl oating on the fruit septum micromorphology are typical of N. micro- water surface near the rush vegetation, on both sides of phyllum and also make it possible to distinguish this the bridge in the village (Fig. ). Analogously as in the species from other representatives of the genus Nastur- two former localities, on October a relevé was tium (Fig. E, F). prepared on a × m plot using the Braun-Blanquet In the discussed locality at an area of × m method. Plants growing along the river were found only a relevé was prepared on August according in layer C (total cover %) and included Phragmites to Braun-Blanquet using a seven-point cover scale and australis ., Carex acutiformis ., Nasturtium micro- a fi ve-point abundance scale. All plants growing both phyllum ., Sagittaria sagittifolia ., Potamogeton na- along and in the river itself were found in layer C (to- tans ., Lemna minor ., Hydocharis morsus-ranae ., tal cover %) and they included Stratiotes aloides ., Typha latifolia .., Berula erecta +, Glyceria maxima +, Nasturtium microphyllum ., Glyceria maxima ., Myosotis palustris +, Nuphar luteum +, Filipendula ulma- Lemna minor ., Phragmites australis ., Berula erecta ria r, Iris pseudacorus r, Lycopus europaeus r, Mentha +, Callitriche stagnalis +, Cirsium oleraceum +, Epilobium aquatica r, Padus avium r, Rumex hydrolapathum r, Salix hirsutum +, Hydrocharis morsus-ranae +, Myosotis palu- triandra r, Scrophularia umbrosa r, Spirodela polyrrhiza r, stris +, Potamogeton natans +, Sium latifolia +, Spirodela Symphytum offi cinale r. The discussed species in local- polyrrhiza +, Urtica dioica +, Veronica anagallis-aquatica ity C is also growing in the Nasturtietum microphylli +, Calystegia sepium r, Epilobium obscurum r, Polygonum community. persicaria r. The analysed species is growing in the de- The genome size of the studied plant material deter- scribed locality in the Nasturtietum microphylli com- mined by fl ow cytometry was . pg/C in octaploid munity classifi ed to the Sparganio-Glycerion fl uviatili N. microphyllum species collected from three localities. association, in the Phragmitetea class. The highest nuclear DNA content was found in plants collected from locality B (‘Rzepin ’), which was . B – locality ‘Rzepin ’ pg/C. A similar C value was obtained for plants from In locality B onerow yellowcress was found in the locality C (‘Starościn’). The lowest C DNA content was Ilanka River on both sides of its m wide bed, over estimated for plants growing in locality A (‘Rzepin ’) a m long segment, in the form of distinct patches and it amounted to . pg (Table ). The statistical fl oating on the water surface near the rush vegetation analysis showed no signifi cant diff erences between all (Fig. ). Only fl owering plants were observed in this lo- these localities. The obtained histograms were of good cality. The relevé for that locality was prepared using the quality (Fig. ) with a low coeffi cient of variation (CV; Braun-Blanquet on October on a × m plot. Table ).

T . Nuclear DNA content of Nasturtium microphyllum leaves collected from three localities near Rzepin. Mean ±SD, n =

DNA content CV (%) CV (%) Locality (pg, mean ±SD) of target species of internal standard

A (‘Rzepin ’) . ±.NS .-..-.

B (‘Rzepin ’) . ±. .-..-.

C (‘Starościn’) . ±. .-..-.

C DNA values between the localities did not show statistically signifi cant diff erences (NS) at P = . (Stu- dent’s t-test). Occurrence of onerow yellowcress Nasturtium microphyllum (Boenn.) Rchb. in the Ilanka River (Lubusz Land)

karyotype of Rosewood (Aniba rosaeodora). Gen. Mol. Biol. : -. C A., M M. (): Występowanie rukwi drobnolistnej Nasturtium microphyllum (Bo- enn.) Rchb. w Wielkopolsce. Chrońmy Przyr. Ojcz. , : -. C A., M M. (): Nowe stanowi- ska rukwi drobnolistnej Nasturtium microphyllum (Boenn.) Rchb. w Polsce. Chrońmy Przyr. Ojcz. – in print. C A., M M., N R. (): Występowanie rukwi płonnej Nasturtium ×sterile w Polsce. Chrońmy Przyr. Ojcz. , : -. D J., B J., V H., G J. (): Nuclear DNA content and genome size of trout and human. Cytometry A: -. H H., M T. (): Bildatlas der Farn- und Blütenpfl anzen Deutschlands. Ulmer Verlag, Stutt- gart. H L., B A., J M.T., B M.D. F. . Cytometric analysis of nuclear DNA content (): First nuclear DNA C-values for eudicot in leaves of Nasturtium microphyllum using Partec buff er families. Ann. Bot. : -. with PI and RNase. Leaves of Petunia hybrida ‘P × Pc’ Internet: http://www.pfaf.org (..). (C = .pg) were used as an internal standard K J., S R. (): Rukiew drobnolistna Nasturtium microphyllum (Boenn.) Rchb. – nowe stanowiska w okolicach Krakowa. Chrońmy Przyr. Ojcz. , : -. Due to the abundance and regular occurrence of the M M., C A., J I. (): investigated species in all the three localities and the Estimation of nuclear DNA content in Nasturtium fact that they are all situated in the vicinity of a large R. Br. by fl ow cytometry. Aquat. Bot. : -. town, a question may arise whether onerow yellow- N A. (): A short note on the genus Na- cress was artifi cially introduced to be more extensively sturtium R. Br. (Cruciferae) and a new hybrid state cultivated. from this genus for Iran. Iran. J. Bot. , : -. P B., P A. (): Polski zielnik ku- linarny. Krajowe zioła jadalne i przyprawowe. Leksy- THREATS AND PROTECTION GUIDELINES kon i przepisy. Wydawnictwo Kurpisz, Poznań. R M Środowiska z dnia stycz- The greatest threat for all the discussed onerow yel- nia r. w sprawie gatunków dziko występujących lowcress populations may be connected with fl oods, roślin objętych ochroną. Dz.U. nr , poz. water pollution and maintenance of river banks and z dnia lipca r. bed. In order to preserve these populations during river R L. (): Klucz do oznaczania roślin na- management works small patches with N. microphyllum czyniowych Polski Niżowej. Wyd. Nauk. PWN, War- would have to be transferred to a substitute water body szawa. and after the completion of river management works S M. (): Nasturtium microphyllum (Boenn.) they would have to be placed back in the Ilanka River. Rchb. In: Polska czerwona księga roślin. Eds R. Kaź- The proposed protection measure is feasible and rela- mierczakowa, K. Zarzycki. Inst. Bot. im. W. Szafera tively easy to perform, since the Ilanka River Valley is a PAN, Inst. Ochr. Przyr. PAN, Kraków: -. protected area within the Natura network (the Ilan- S J., K T., F R. (): Nuclear DNA ka River Valley – Dolina Ilanki, area code: PLH ). amounts in Macaronesian angiosperms. Ann. Bot. : -. Ś E., Z E., J I. (): Are REFERENCES seeds suitable for fl ow cytometric estimation of plant genome size? Cytometry A: -. B W., H H., K M. (): Zum Vorkom- T T. (): Nasturtium R. Br., Rukiew [Water cress]. men und zur Morphologie von Nasturtium sterile In: Flora Polski. Ed. A. Jasiewicz. Vol. . Rośliny na- (Airy Shaw) Oef. in Südwestniedersachsen und an- czyniowe. PWN, Warszawa–Kraków: -. grenzenden Gebieten. Flor. Rundb. , : -. B W., H M., H H. (): Evoluti- on of hybrid taxa in Nasturtium R. Br. (Brassicaceae). For citation: Czarna A., Morozowska M., Jędrzejczyk I. Folia Geobot. : -. (): Occurrence of onerow yellowcress Nasturtium C L.A.S., C C.R., M F.A., microphyllum (Boenn.) Rchb. in the Ilanka River (Lubusz F D.V. (): Nuclear DNA content and Land). Rocz. AR Pozn. , Bot. Stec. : -.