Some interesting records of Cladonia species from the Nizina Wielkopolska Lowland (W Poland)

The lichen genus Cladonia comprises several similar species which have hardly been recognized in Western Poland so far. We used thin layer chromatography (TLC) as a simple technique to determine diagnostic lichen substances in morphologically similar Cladonia species. During field studies in Sandr Nowotomyski (western Nizina Wielkopolska Lowland), ten interesting records of Cladonia species were made. Cladonia novochlorophaea is reported for the first time from this region. Our records supplement the knowledge about the distribution of Cladonia species both in the investigated region and in Poland. All the records are compared with existing literature data from regional floristic inventories and distribution maps from Poland.


INTRODUCTION
The lichen genus Cladonia Hill ex P. Browne comprises several common and widely distributed epigeic species, well known as reindeer or cup lichens. Detailed chemical investigations in the 20 th century revealed that secondary lichen compounds show a high diversity in cup lichens, and several new species were described on the basis of these lichen substances (e.g., Hammer 1995). Recently, the importance of secondary lichen metabolites to the taxonomy and species discrimination within the genus Cladonia has been confirmed in a first molecular approach by Stenroos et al. (2002). Though Cladonia species can be found in many lichenological reports from different regions of Poland (Fałtynowicz 2003 and literature cited therein), the information about the species recognizable only by secondary substances is rather ACTA MYCOLOGICA Vol. 44 (2): 223-232 2009 Dedicated to Professor Krystyna Czyżewska in honour of 40 years of her scientific activity sparse. Microcrystallization, thin layer chromatography (TLC), or high pressure liquid chromatography (HPLC) are often the only methods to determine specific secondary metabolites (Orange et al. 2001), but in Poland only a few studies have used these laboratory techniques for the identification of Cladonia species so far (e.g., Kowalewska et al. 2000Kowalewska et al. , 2008Kowalewska, Kukwa 2003Kowalewska, Szok 2004;Kukwa 2005a, b;Osyczka 2006;Oset et al. 2008;Syrek, Kukwa 2008).
Recent lichen inventories carried out in Sandr Nowotomyski in the western part of Wielkopolska lowland in the years 2007 and 2008 revealed several stands of rare and noteworthy Cladonia species. Some taxa have already been known from the central part of the area in the Nowy Tomyśl region (Krawiec 1938;Dziabaszewski 1962;Nowacka-Zyber 1967;Żukiel 1967;Bujakiewicz, Lisiewska 2003;Zarabska 2008a, b). We restrict our study to rare Cladonia species and those which require TLC analysis for the identification. Our data supplement the information about the distribution range in Poland for the following species: Cladonia cariosa, C. chlorophaea s.str, C. coccifera, C. crispata, C. grayi, C. merochlorophaea, C. novochlorophaea, C. pyxidata s.str., C. rei and C. subulata.

MATERIAL AND METHODS
Specimens collected during field work were identified with the help of Nowak and Tobolewski (1975), Purvis and James (1992), and Wirth (1995). For the analysis of morphologically similar Cladonia species, thin layer chromatography was performed in solvents A and C according to the standard method of Culberson and Ammann (1979). Nomenclature follows Fałtynowicz (2003). All samples were collected by the first author, and the specimens are kept in the Natural History Collection UAM Poznań (POZ).
In the following list of species we give precise information about location, detected secondary compounds (if TLC was carried out), about earlier notes in the literature, and about habitats.

Cladonia grayi
Cup lichen with gradually tapering, regular, trumpet-or goblet-shaped scyphus, often with proliferations from cup margin. In the upper part, the podetia are covered by granular soredia, while in the lower part, the stalk is corticated, distinctly verruculose and occasionally squamulose (Kowalewska et al. 2008). According to Kowalewska et al. (2008), Cladonia grayi is almost absent in Western Poland and partially also in Eastern Poland; from the Nizina Wielkopolska Lowland it has only been reported by Kubiak (2008) so far. In the investigated area the species was found in a clearing of birch and pine forests.

Cladonia merochlorophaea
Characterized by tall, brownish, greenish-grey podetia with gradually fl ar-haracterized by tall, brownish, greenish-grey podetia with gradually flaring scyphi and areolate-corticate, verruculose, or obscurely sorediate to coarsely granular surface (Kowalewska et al. 2008), morphologically indistinguishable from C. novochlorophaea. Several localities of C. merochlorophaea were discovered in the western Poland by Kowalewska et al. (2008). In Wielkopolska it was previously noted only by Rakowski (2003) and Zarabska (2008a, b), but seems to be the most common member of the Cladonia chlorophaea-group in this region. In accordance with results of Kowalewska et al. (2008), C. merochlorophaea was mainly recorded on humus-rich soil.
Substances detected by TLC: merochlorophaeic and 4-O-methylcryptochlorophaeic acid (present in all specimens), furmarprotocetraric acid (present in four of seven studied specimens). According to Kowalewska et al. (2008), this cup lichen is morphologically indistinguishable from C. merochlorophaea and was only recently discovered in Poland (Kowalewska, Kukwa 2001). Until now, it was rarely noted in the country and was recorded mainly from the northern part (Kowalewska et al. 2008). It can be distinguished from C. merochlorophaea by the presence of homosekikaic and sekikaic acids. Here, the species is reported for the first time for the Nizina Wielkopolska Lowland.

Cladonia rei
Characterized by long, granular-sorediate to farinose podetia with or without narrow cups with short proliferations, by often prominent apothecia, or by stout podetia with short anisodiametric branching patterns. This species can be confused with the morphologically similar C. glauca and C. subulata (Kukwa 2005a). In Poland, Cladonia rei is common in the eastern part of the country and has been recorded from the Wielkopolska Lowland only once (Syrek, Kukwa 2008).
Lately, the taxonomical status of C. rei has been under discussion (Spier, Aptroot 2007;Syrek, Kukwa 2008); however, most recent molecular studies by us (Dolnik, Beck and Zarabska subm.) proved the distinctness of C. rei as a separate species, which is not closely related to C. subulata.
Substances detected by TLC: homosekikaic acid (present in all specimens), furmarprotocetraric acid (present in two of four studied specimens). Characterized by long subulate podetia with a granular to farinose sorediate sur-haracterized by long subulate podetia with a granular to farinose sorediate surface, often with an antler-like branching pattern, and by prominent long-stalked cups with long and unequal proliferations narrowing to an acute tip. In contrast to Cladonia rei, this species was often recorded in the Wielkopolska area (Krawiec 1930(Krawiec , 1933(Krawiec , 1938(Krawiec , 1955Tobolewska, Wronówna 1955;Dziabaszewski 1962;Tobolewski 1963;Glanc 1969Glanc , 1998Glanc et al. 1971;Kozłowska 1975;Tobolewski, Kupczyk 1977;Balcerkiewicz, Brzeg 1993;Brzeg, Pawlak 1998;Kepel 1996Kepel , 1999Rakowski 2003), but misidentifications through confusion with furmarprotocetraric acid containing morphs of C. rei cannot be ruled out completely. Cladonia subulata was recorded in six new localities in the western part of Wielkopolska. These were also the first recordings of this species for Sandr Nowotomyski.
Substances detected by TLC: furmarprotocetraric acid (present in all specimens).

CONCLUSION
Western Poland, especially sandy areas with inland dunes, comprises an interesting epigeic lichen flora with several Cladonia species. The use of TLC for the identification of lichens collected in the Wielkopolska region revealed several records of rare species, which have been absent in older sources of Poland. The previously poorly recognized species Cladonia merochlorophaea turned out to be the most common member of the Cladonia chlorophaea-group in the region.