ACTA BOTANICA UNIVERSITATIS COMENIANAE

Volume 46

2011 COMENIUS UNIVERSITY IN BRATISLAVA

The journal was edited with the title / Časopis bol vydávaný pod názvom Acta Facultatis Rerum Naturalium Universitatis Comenianae, Botanica

Editor in Chief / Predseda redakčnej radyKarol Mičieta; micieta@fns.uniba.sk

Executive Editor / Výkonný redaktorSoňa Jančovičová, jancovicova@fns.uniba.sk

Editorial Board / Členovia redakčnej radyDana Bernátová, Danica Černušáková, Zuzana Dúbravcová, Katarína Mišíková

Editor Ship / Adresa redakcieRedakcia Acta Botanica, Révová 39, SK-811 02 Bratislava 1; Tel. ++421 2 54411541; Fax ++421 2 54415603

Published by / Vydavateľ© Comenius University in Bratislava, 2011© Univerzita Komenského v Bratislave, 2011

ISBN 978-80-223-3113-5ISSN 0524-23712

BIOTRANSFORMATION OF SOME GLYCOSIDES BY ß-GALACTOSIDASE IN IMMOBILIZED CELLS OF DIGITALIS LANATA

Karol Mičieta1•, Ján Stano2, Marcela Koreňová3, Beate Dietrich4, Magdaléna Valšíková5

1Comenius University in Bratislava, Faculty of Natural Sciences, Department of Botany, Révová 39, 811 02 Bratislava, Slovakia

2Comenius University in Bratislava, Faculty of Pharmacy, Department of Cell and Molecular Biology

of Drugs, Odbojárov 10, 832 32 Bratislava, Slovakia3Comenius University in Bratislava, Faculty of Pharmacy, Garden of Medicinal Plants, Odbojárov 10,

832 32 Bratislava, Slovakia4Martin Luther University, Institute of Pharmaceutical Biology and Pharmacology, Hoher Weg 8,

061 20 Halle/Saale, Germany5Slovak University of Agriculture in Nitra, Horticulture and Landscape Engineering Faculty, Department

of Vegetables-Production, Trieda A. Hlinku 2, 949 76 Nitra, Slovakia

Received 5 July 2011; Received in revised form 9 August; Accepted 16 September 2011

Dedicated to Professor Jozef Tomko and Associated Professor Jaroslav Kresánek, on the occasionof their 90th birthdays.

Abstract

The cells of Digitalis lanata Ehrh. (foxglove) were after permeabilization with Tween, ethanol,hexadecyltrimethylammonium bromide and hexadecylpyridinium chloride resp., immobilized by glutaraldehydewithout any soluble carrier. β-Galactosidase showed pH optimum at 4.8 and temperature optimum at 50 °C. Theenzyme hydrolysis was linear for 3.5 h reaching 60 % conversion. Cells immobilized by cross-linking having highβ-galactosidase activity and a long-term storage stability showed convenient (appropriate) physico-mechanicalproperties.

Key Words: Digitalis lanata Ehrh., β-galactosidase, cell permeabilization, cell immobilization

Introduction

Sources of numerous natural compounds are limited. Synthetic preparation of these compoundscomplements their insufficiency which is being limited by natural sources. Solution of this problem isaided by biotechnologies. Biotransformation and production of high-value fine and special chemicalsare known from recent time. Knowledge of totipotency and mastering of plant-tissue cultivationtechniques were applied at first in agriculture for instance in plant propagation. It was recognized laterthat plant cells can be used for biosynthesis and biotransformation of various substances of natural andsynthetic origin.

In the last decades several methods for fixation of biocatalysts have been developed. Enzymes,living or nonliving microorganisms, animal and plant cells, as well as combined systems, have beenbound within or to carrier materials (Hulst, Tramper 1989; Förster 1994; Klein, Wagner 1983; Rosevear1984; Akin 1978). Immobilization of cells or enzymes represents an effective way of highly efficient

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Acta Botanica Universitatis Comenianae Vol. 46, 2011

• Corresponding Author: Karol Mičieta; micieta@fns.uniba.sk

enzyme fixation of catalysts important for biotransformation process (Blandino et al. 2003). Plant cellswere first immobilized by Brodelius et al. (1979). Many matrices from synthetic polymers or biologicalmaterials have been used for the immobilization of cells. The most widely used technique of cellimmobilization is cell entrapment when living cells are enclosed into gel particles: agar, agarose,kappa-carrageenan, collagen, chitosan, polyacrylamide, polyurethane, or cellulose (Hulst, Tramper1989; Tampion, Tampion 1987). The spontaneous adhesion or covalent binding of cells to the surfaceof insoluble carriers was also examined (Cabral et al. 1984, Parascandola et al. 1987, Szczodrak 1999).Recently, the use of poly(vinyl alcohol) and glutaraldehyde (Wu, Wisecarver 1992; Hasal et al. 1992)or Tween 80 and glutaraldehyde (Barth et al. 2002) for cell immobilization have been investigated.

Biotransformation of foreign substrate into more useful substances by cultured plant cells is one ofthe important reactions in synthetic chemistry (Sakamaki 2001). From the viewpoint of “greenchemistry”, biocatalysts have been used for the transformation of natural or artifitial substances intouseful compounds (Flitsch 2002) by epoxidation (Sakamaki 2008), oxidation or oxidative cleveage(Chai 2004, Itoh 2008, Utsukihara et al. 2007) reduction (Itoh 2005) conversion (Sakamaki 2004,2005). Glycosylation with plant cells is also an important method for the conversion of water insolubleand unstable compounds in water-soluble and stable once. Cultured plant cells showed high potentialfor glycosylation phenolic compounds to afford the corresponding mono- and disaccharides (Shimoda2006, 2007a, b, 2009). Biological degradation of Bisphenol A was studied by cultured plant cells. Chaiet al. (2003) reported the total degradation of Bisphenol A in aqueous solution by using Caraganachamlangu in biocatalytic reaction. For biotransformation of some bioproducts of plant origin can beuseful also microorganisms (Keshk 2011).

β-galactosidase (β-D-galactoside galactohydrolase EC 3.2.1.23) called also lactase catalyses thehydrolysis of terminal β-galactosidic linkage of glycosides. The enzyme is widely distributed in variousplant tissues. It is involved in the degradation of plant cell wall polysaccharides in relation to cellgrowth, rippening of fruit and seeds, and pollens germination (Simon et al. 1979; De Veau et al. 1993;Sawicka, Kacperska 1995; Singh, Knox 1985). Lactase hydrolyses lactose into glucose and galactosewhich has recently evoked considerable interest because of its application in the food industry, nutritionand medicine (Szczodrak 1999, Tatsuzawa et al. 2003). Although lactase is generally present in plants,this source has been generally neglected.

The enzymatic hydrolysis of terminal β-galactosidic linkage of glycosides by cells, as well as byfoxglove cells immobilized has been studied in this paper.

Material and Methods

Plant material. – Long-term callus culture and cell suspensions were derived from foxgloveseedlings (Digitalis lanata Ehrh.) as was described previously (Diettrich et al. 1986). Seedlings offoxglove grew under sterile condition 5-6 days (Blanáriková et al. 1996, Dixon, 1991).

Cell permeabilization. – Cell suspensions were filtered through a nylon cloth, and 10 g of freshmass was suspended in 50 ml of 0.15 mol.l-1 NaCl with 5 % Tween 20, 5 % Tween 80, 30 % ethanol,50 % ethanol, 0.1 % hexadecyltrimethylammonium bromide and/or 0.1 % hexadecylpyridiniumchloride, respectively. Permeabilization was carried out for 3 h with moderate stirring at 20 °C. Thecells were filtered off and washed with 2000 ml distilled water and 3000 ml 0.15 mol.l-1 NaCl solution.

Cell immobilization. – The permeabilized cells were immediately suspended in 40 ml of 0.15mol.l-1 NaCl solution and than 5 ml of 25 % glutaraldehyde was added slowly under mild stirring for2.5 h at room temperature. The immobilized cells were then separated and washed with 2000 ml ofdistilled water and 2500 ml of 0.15 mol.l-1 NaCl solution.

Determination of fresh and dry weight. – Fresh and dry weight of suspended cells andimmobilized cells were determined gravimetrically. For determination of dry weight, samples weredried to the constant weight at 100 °C.

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Glucose utilization. – The immobilized cells and cell suspension were exposed to initial glucoseconcentration (200 mg.l-1) in the cultivation medium (Yagi 2004) without the presence of sucrose. The concentration of glucose was determined by the method of Trinder (1969).

Enzyme assay. – The enzyme assay was performed by a modified method of Kim et al. (2002)using p-nitrophenyl-β-D-galactopyranoside (β-PNG) as substrate. The reaction mixture contained 0.1 g of fresh cells and 0.5 mg of β-PNG in 2 ml Mc Ilvaine buffer, pH 4.8. The control containedboiled cells. Both mixtures were kept in a period of 20 min to 5 h at 30 °C using a rotatory shaker (80 rpm) and the reaction was stopped by 2 ml of 1 mol.l-1 Na2CO3. The p-nitrophenol released wasdetermined spectrophotometrically at 420 nm. Cells were separated from the reaction mixture, driedand enzyme activity was calculated per 1 g of dry mass (Barth et al. 2002). The enzyme activity wasexpressed in katals. Protein content was determined by the method of Doumas et al. (1981) usingbovine serum albumin as the standard.

Storage stability. – The stability of melibiase during storage was monitored in the followingexperiments. The immobilized cells were storied at 4 °C in 0.15 mol.l-1 NaCl with addition of thefollowing compounds: a) chloramphenicol 50 mg.l-1, b) chlortetracycline hydrochloride (CLCTC) 50 mg.l-1, c) (1-methyldodecyl)dimethylamine N-oxide (ATDNO) 100 mg.l-1 (Devínsky et al. 1978).

Cell viability. – Cell viability was determined by Dixon (1991) with 2,3,5-triphenyltetrazoliumchloride (TTC), fluorescein diacetate and with an oxygen electrode, respectively.

Results and Discussion

Glutaraldehyde immobilized foxglove cells showed little morphological changes in comparisonwith cells in suspension. A small cell plasmolysis and some aggregation of the cells were occurringduring immobilization. The cells immobilized with glutaraldehyde did neither consume glucose (Fig. 1) nor were viable, since they did not, show any respiratory activity and were not stained withfluoresceindiacetate or 2,3,5-triphenyltetrazolium chloride.

The permeabilization of the studied foxglove cells by Tween 20, Tween 80, hexadecylpyridiniumchloride (HPCH) and hexadecyl(trimethyl)ammonium bromide (HTAB) led to the leakage ordegradation of proteins. Enzyme activity showed a moderate increase. By glutaraldehyde cross-linkingonly a moderate increase in the enzyme activity has been found (Tab. 1).

Sucrose is probably the most widely used carbon source in plant tissue cultures. The utilization ofsucrose by foxglove cell culture is followed by a consistent pattern of rapid initial inversion andsequential phases of glucose and fructose consumption. Glucose and fructose are present in the mediumin roughly equal amounts after the first few days of inoculation, but the cells did not consume fructoseuntil glucose is present (Hamilton et al. 1984). Figure 1 shows the results of glucose utilization by cellsuspensions and by glutaraldehyde cross-linked cells.

The pH optimum of lactase in immobilized cells is at 4.8 like in viable cells. Enzyme hydrolysis ofβ-PNG was linear for 3.5 h reaching 55–60 % of conversion and after that time it practically stops. Thetemperature optimum for immobilized cells and free cells was at 50 °C. Similar properties werereported for lactase isolated from winter rape (Sawicka, Kacperska, 1995). The inhibitory effect of 0.1–0.5.10-3 mol.l-1 4-(chloromercurio)benzoic acid can be eliminated by 5–10.10-3 mol.l-1 cysteine,dithiothreitol, or by 2-mercaptoethanol, respectively (Barth et al. 2002, Budík 1992). These resultsindicate that -SH groups are essential for the activity of α- and β-galactosidase (Budík 1992, Machová1994).

Partially purified enzyme preparations of lactase from gherkin and poppy seedlings were inhibitedby galactose and glucose in a moderate way (Budík 1992, Machová 1994). A similar inhibitory effectwas observed in immobilized cells. As illustrated in Tab. 2, the activity of this enzyme in foxglove cellsimmobilized by glutaraldehyde (in 0.15 mol.l-1 NaCl with all preservatives) during 6 months storage isstill relatively high. It is known, that immobilization of plant cells by entrapment in beds compared

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with free cells brings some important advantages. It encourages release of enzyme reaction products,prevents cell aggregation, protects cells from shear stress, gives good cell-to-cell contact, and it preserves the activity of multifunctional systems (Rosevear, 1984). The formation of cell aggregatesin cell suspension and the degree of cell differentiation are important from the biotechnological pointof view. In cells immobilized by cross-linking with bifunctional reagents (e.g. glutaraldehyde), theactivity of tyrosine decarboxylase, DOPA-decarboxylase and α- and β-galactosidase (Blandino et al.2003, Barth et al. 2002, Machová, 1994, Stano et al. 2002) still reach high values.

Glutaraldehyde is therefore successfully used for immobilization of poppy, carrot, gherkin,california poppy and other cells of plant origin (Barth et al. 2002, Stano et al. 2002).

The immobilization costs are very low and no special equipment is needed. Immobilization ofwhole cells makes enzyme isolation unnecessary, whereas the enzyme activity remains high. Both thereaction kinetic parameters and the physico-mechanical properties of the biocatalysts are fullycomparable to those of biocatalysts prepared by immobilization (Hasal et al. 1992).

Immobilized lactase and other glycosidases can be applied in biotransformation processes ofpharmaceutically important compounds as well as in food industry (Mučaji et al. 2001, Tatsuzawa et al. 2003, Nunez et al. 2003, Bezáková et al. 2007). The structural study of the compounds is anotherpossible field of their practical use (Gupta et al. 2003, Munkombwe 2003, De León et al. 2003,Holková et al. 2011).

Tab. 1. Protein content of β-galactosidase activity in 10 days foxglove suspensions. Permeabilized by 5 %Tween 20, 5 % Tween 80, 0.1 % hexadecyl(trimethyl)ammonium bromide (HTAB) and 0.1 %hexadecylpyridinium chloride (HPCH) and cells immobilized by glutaraldehyde. The values are expressedas means of five replicates (n = 5)±SDE. For details, see Exper. Part.

Tab. 2. Stability of β-galactosidase in immobilized foxglove on storage

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Cells Protein Activity Specific activity(mg/g dry mass) (nkat/1 g dry mass) (nkat/mg protein)

Suspension 14.3±0.09 7.2±0.10 0.50

Permeabilized 0.1% HTAB 6.8±0.09 5.8±0.09 0.85

0.1% HPCH 6.8±0.09 5.7±0.09 0.84

5% Tween 20 6.8±0.08 5.8±0.10 0.85

5% Tween 80 6.8±0.08 5.8±0.09 0.85

Immobilized 0.1% HTAB 6.7±0.09 5.7±0.10 0.85

0.1% HPCH 6.7±0.09 5.6±0.09 0.84

5% Tween 6.7±0.08 5.6±0.09 0.84

5% Tween 6.7±0.08 5.6±0.10 0.84

Conservance Original activity in suspension culture (% month)

0 1 2 3 6

None 100

CLCTC (50 mg/l)65 67 69 75 85

ATDNO (100 mg/l) 65 67 69 75 86

Chloramphenicol (50mg/l) 63 67 70 76 88

Sodium azide (200 mg/l) 63 66 70 76 89

Frozen in 0.15 M Nacl 63 66 71 77 90

0

100

65

65

63

63

63

1

67

67

67

66

66

2

69

69

70

70

71

3

75

75

76

76

77

6

85

86

88

89

90

Fig. 1. Time course of glucose consumption in foxglove cells immobilized with glutaraldehydeand in cell suspension

Acknowledgements

The research was supported by the VEGA grant Agency (Bratislava) grant No. 1/0182/09 andpartially by the Research project of the Ministry of Agriculture “Development of progressive methodspractices for continuous quality improvement in the process of food production and monitoring”.

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Abstrakt

Bunky suspenznej kultúry Digitalis lanata Ehrh. (náprstník vlnatý) sa po permeabilizácii Tweenom,etanolom, hexadecyltrimetylamonium bromidom a hexadecylpyridinium chloridom imobilizovaliglutaraldehydom bez nosiča. Enzýmová hydrolýza má lineárny priebeh počas 3,5 h a dosahuje 60 %konverzie. β-galaktozidáza v imobilizovaných bunkách má pH optimum pri 4,8 a tepelné optimum pri50 °C Imobilizované bunky majú vysokú enzýmovú aktivitu, dlhodobú stabilitu a vhodné mechanickévlastnosti.

Karol Mičieta, Ján Stano, Marcela Koreňová, Beate Dietrich, Magdaléna Valšíková: Biotrans -formácia vybraných glykozidov β-galaktozidázou v imobilizovaných bunkách Digitalis lanata.

9

APPLICATION OF AGING THEORIES TO PLANT ORGANISMS WITH AN EMPHASIS ON TREES

Eva Brutovská•, Karol Mièieta, Andrea Sámelová

Comenius University in Bratislava, Faculty of Natural Sciences, Department of Botany, Révová 39, 811 02 Bratislava, Slovakia

Received 2 August 2011; Received in revised form 15 September; Accepted 4 October 2011

Abstract

The main question of aging theories is, if aging is programmed or not, and how did it evolve. While these questionshave not yet been clearly resolved, several groups of possible theories have been published on this topic. However,most of these theories do not consider plants and their specific traits, making the mechanisms of their aging unique.The first trait is clonality and sectoriality, the second is the lack of a differentiated germ line. The lack of germ lineprevents telomere shortening, and can lead to the transfer of somatic mutations into sexual offspring, whilesectoriality in trees causes the isolation of potentially catastrophic events into one part of the tree, making it a population of more or less independent modules within one axis. The processes of population dynamics, includingaging, can act with different impact within the frame of population as within the frame of an individual tree as well.

Key words: aging, plant senescence, dendrology

Introduction

Aging is defined as a complex of age-related changes in the organism, increasing the probability of its death (Khohklov 2010). If the process of extinction in the population follows an exponential lawi.e. with a constant rate of mortality, we can consider the aging of this organism negligible, irrespectiveof their actual lifespan. The basic question of aging theories is, if this process is programmed or not.The term “program” is defined as a sequence of coded instructions that is a part of an organism. In theterms of this definition, apoptosis can be considered programmed cell death only in the case, when it occurs during the embryonic development as the elimination of cells that are no longer necessary forthe development of the organ or tissue. An aging program would be therefore a program, whichincreases the probability of death of the organism in dependance on its age (Khohklov 2010).

Aging is a universal principle for all living organisms, but is not a biologically inevitable process– living systems are able to maintain immortality at the cell level (Heininger 2002). Aging is favouredby natural selection, and many authors suppose it’s genetically programmed (Finch 1990, Rose 1991,Finch, Ruvkun 2001). The standard arguments of programmed aging are: (1) different characteristiclifespan of individual species, (2) heritable lifespan, (3) the existence of progeroid diseases, (4) increasein the lifespan of experimental animals after the knock-out of certain genes, (5) existence ofprogrammed death in some organisms. However, against each of these facts, there are the arguments ofthe “antiprogramists” (Khokhlov 2010): The different lifespan of individual species can be explainednot by different aging programmes, but different development programmes, which create organismswith different reliability of functions – just like in the case of cars or electronics, which also have noaging programme, and despite it the probability of unrepairable damage increases with their age, anddepends on the quality of the product. Therefore is the lifespan also heritable and progeroid diseases

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Acta Botanica Universitatis Comenianae Vol. 46, 2011

• Corresponding Author: Eva Brutovská; stromosestra@gmail.com

don’t cause the acceleration of aging programme, but lowering the organism’s defences against theinfluences, which cause ageing. The increase in the lifespan of experimental animals after the knock-out of certain genes (like age-1 and daf-2 in Coenorhabditis elegans) is also ambiguous. Some of thedaf-2 mutants die in the age of 10 days, while others in the age of 50 days (in comparison to the 2-3weeks lifespan of the wild type). Moreover, these mutants are weaker in the terms of viability when putinto competition with the wild type (Jenkins et al. 2004). The programmed death cannot be identifiedwith programmed aging, as its probability does not increase with age, as it results from the definitionof aging. The main feature of any programme is, that there is a possibility of its violation. It is valid forthe development programme, but there has been no evidence about a violation of the aging programme,resulting in an “immortal” individual.

Between the arguments of programists and anti-programists, no consistent theory of aging has beenelaborated so far. Several groups of theories have been proposed, explaining the process of aging witha different relation to the evolution theory and population genetics.

When speaking about plant aging or senescence, it is necessary to define the terms first. Munné-Bosch (2008) discerns two different conceptions of the term. According to the first conception,senescence is a highly regulated process on molecular, biochemical and physiological niveau, leading tothe death of cell, tissue, organ or entire plant. It represents the last ontogenetical stadium in the life ofthe plant. Aging, on the other side, is a degenerative, passive process, which is not endogenouslyregulated (Noodén 1988). Van Doorn and Joshimoto (2010) defined aging as a deterioration process,which increases the probability of death with the increasing chronological age, and is used to label anychanges connected with age. The term senescence concerns not only an individual, but also the processespreceding the necrosis of organs in an otherwise healthy individual, which can be beneficial to theorganism. Therefore they recommend using the term “whole plant senescence” to describe the processequivalent to the aging of animals. Thomas (2002) defined senescence as a physiological state precedingdeath in the majority of cells, tissues and organs. It is the way of reformation the plant by the removalof unwanted or inconvenient cells and tissues with a simultaneous relocation of resources, precedingaging. Senescence is mechanically independent of degeneration and death, and doesn’t have to lead toit, neither. In the case of many species senescence is even reversible (Zavaleta-Mancera et al. 1999).

Šebánek (1998) maintains the opinion that the aging of cells, tissues, and whole plants isprogrammed, as it results from the study of gene expression during the process of aging. Thisdefinition, however, is closer to the definition of senescence. The symptoms are observable in all cellparts: structural changes of DNA, chromosome aberrations, the speed of RNA and protein degradationincreases, as well as the synthesis of protheases, ribonucleases and chlorophyll degrading enzymes.Degeneration of chloroplasts, endoplasmatic reticulum and tonoplast occurs, later also mitochondrias,nucleus and plasmalema. The rate of photosynthesis decreases, while the respiration rate increases.Free amino-acids are translocated to the growing parts of the plant (Šebánek 1998).

According to the second conception (Charlesworth 1980) senescence is understood from the pointof view of population biology as the decrease of age-specific value of survival and fertility, which alsodecreases the performance of many physiologic functions. This conception is similar to agingaccording to the previous theory, but concerns only later ontogenetic stages. Vaupel et al. (2003) haveeven proved the possibility of negative senescence in population models – the case when mortalitydecreases with age after reaching the sexual maturity.

When speaking about plant senescence, Munné-Bosch (2008) states significant differences betweenmonocarpic and polycarpic plants. In the life history of monocarpic plants two kinds of senescence canbe observed: the first occurs during vegetative growth by remobilisation of nutrients from older parts ofthe plant to the younger ones. The second kind concerns all organs, and leads to the death of the wholeplant by resources remobilisation into the seeds. In the polycarpic plants, the repeated investment ofresources into reproduction can lead to a decrease of survival rates, but the aging is gradual, and can beassociated with a remarkable life span. It can be concluded, that senescence according to the firstconception – as a controlled endogenous process – does probably not occur in polycarpic plants, whilethe second conception – decreasing the survival rates with age – does. However, it has been proved that

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the mortality rates in the populations of many plant species from plankton to trees develop in such a way,that they correspond with the natality rates (Marbà et al. 2007). The only potential proof to support thisfact would be the discovery of a genetic program of aging in plants.

Munné-Bosch (2008) raised the question, if there are differences in the mechanisms of senescencebetween monocarpic and polycarpic plants also on the level of organs. Senescence occurs here asendogenously regulated process, an example of which is the falling of the leaves of the trees in autumn.This event is accompanied by changes in gene expression and the transition from photosynthesis to theenergy production by mitochondrial respiration, oxidation of fat acids and remobilisation of resources.This process is similar like in monocarpic species, but differs by certain factors. In monocarpic speciesduring the lack of water and nourishment senescence begins from the oldest leaves, and in the case ofa long-term stress continues to the youngest parts of the plant with the goal of finishing the life cycleand producing seeds. In polycarpic plants only the oldest leaves wither, and also in the case of a strongstress which induces the senescence of young leaves the plant strives to keep viable meristemes whichwould ensure growth after the stress period.

We can conclude that whole plant senescence has two different forms – monocarpic and polycarpic.The study of Van Doorn and Joshimoto (2010) suggests, that chloroplasts have at least a passive rolein monocapric senescence, and support some symptoms preceding death actively. The knock-out of thechloroplast gene ndhF led to a considerable delay of leaf necrosis after the formation of flowers (Zapataet al. 2005).

According to Larson (2001), when considering plant aging and senescence, it is necessary to discernbetween a genet and ramet. In unitary organisms that reproduce sexually, the individuals continue in theirexistence in the following generations by the means of gametes. Senescence concerns the individualorganisms producing the gametes. Plants represent modular organisms, which colonize time and space bytwo means: genets, as independent, genetically separated entities and by the mitotic repetition of the bodyabove or below ground by ramets – not reproduction, but expansion. When considering the maximum ageof an organism, the age of a genet and ramet is often mistaken. If we consider the age of a ramet, thenthere are species, which have been reproducing only mitotically for millions of years.

The currently formulated aging theories can be divided into several groups. They have been mostlybased on the characteristics of animals. This contribution deals with their possible implications andspecifics with regards on the plant organisms.

1. Theories of damage accumulation

1.1. General theories of oxidative damage accumulationThese theories are based on the accumulation of damage in living structures with age, and the high

price of reparation. Hamilton (1966) supposed that perfect reparation of tissue is not possible; thereforeaging is a biologic necessity. There are two main types of damage, considered in these theories –oxidative damage of biomolecules caused by reactive forms of oxygen, and DNA damage.

The damage by reactive oxygen forms is one of the most recognized causes of aging (Harman1956), supported by the evidence about increasing level of oxidation of DNA, proteins and lipids withage. The most significant source of reactive oxygen forms in the cell is the activity of mitochondrialrespiratory chain, mostly complexes I and II (Skulachev et al. 2009). Lambert et al. (2007) observedthat the lifespan of birds and mammals increases with the decrease in the rate of H2O2 generation in thereverse transport of electrons in the complex I in the mitochondria of the heart. This suggests, that theprotection of cells before intramitochondrial reactive oxygen forms can lower the rate of aging. Naturalantioxidants, like the E vitamin, are degraded by enzymatic processes in the cell in excess, which canbe an indication that aging is programmed, and the organism is trying to perform the program(Skulachev et al. 2009).

The argument against this theory is, that continuous reparation is energetically more favourablethan building a new organism from the zygote. Therefore this theory is not sufficient for explainingaging from the evolutionary point of view (Mitteldort, 2010). Despite that, there is evidence, that

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reparation processes are progressively suppressed in aging individuals. The concentration of enzymes,which degrade the reactive side products of the Krebs cycle, has a decreasing tendency with age inhumans (Tatone et al. 2006). Similar evidence exists about the reparation mechanisms of DNA (Engelset al. 2007). The decreasing activity of protection mechanisms with age, which enables theaccumulation of damage, is one of the primary mechanisms of aging that should be explained (Sanz et al. 2006).

In plants, the theory of damage accumulation has its specifics, especially when it comes to thewoody plants. According to the theory of Thomas (2002), trees avoid senescence by the strategy of“building on the ruins” – creating new, young structures on the accumulated remnants of previousgenerations of tissues and organs and maintaining the genetic integrity in cell lines. Endogenoussenescence can be excluded from the reasons of mortality of trees (Larson, 2001). Therefore onlypathogens and physical damage remain – which can be viewed as damage accumulation with age aswell. Long-lived trees have mostly very high resistance or tolerance to pathogens due to the highdensity and chemical composition of the wood. In short-lived trees the pathogens trigger a wide scaleof defence mechanisms, which moderate their impact until the costs reach the level of the carryingcapacity. The netto primary production, and therefore also the ability to accumulate resources,including those for the fight against pathogens, decreases with age. Ryan and Yoder (1997) however,accredit this rather to the tree size than to its age. The main reason of death for the long-lived trees,especially conifers, is physical damage (Larson 2001).

1.2. Oxidative damage accumulation in plantsOxidation damage occurs in plants as well: especially the covering tissues in a direct contact with

the outer environment and high metabolic activity, like the cork (phelem), are exposed to it. Cork is theouter layer of a covering tissue, consisting of several layers of cells, which deposit large quantity of suberin, and undergo programmed cell death. This tissue is also exposed to a great oxidation stressdue to the fenoxy radicals generated during the suberin synthesis (Whetten, Sederoff 1995). Pla et al.(2000) studied the frequency of mutations in the cork and in root tips of Quercus suber by analysingthe Qs_hsp17 sequences in cDNA and genomic DNA. Qs_hsp17 codes small heat-shock proteins of thecytoplasmatic class I, which are substantially over-expressed in the cork, and other cells with highoxidation stress, as well as in the apical meristhemes. In other tissues, their expression can be inducedby different stressors, including oxidation stress. Pla et al. (2000) found a significantly higher mutationrate in the cork DNA than in the DNA of root tips, especially the types of mutations associated withoxidation damage. That implies, that the genetic integrity is not maintained in a senescent tissue,exposed to oxidation damage. The strategy of cell death intermediated by hypermutation of genes inthe tissue bordering with external environment can help to protect the deeper lying tissues, and so leadto a selection advantage.

1.3. General theories of mutation accumulation The first to formulate this theory was Medawar (1952) when he expressed the theoretic assumption

that the selection pressure lowers with age. Later Edney and Gill (1968) stated, that if the selectionpressure is really close to zero, then the mutation load itself should be a sufficient explanation for theoccurrence of aging in evolution.

The DNA is exposed to a steady pressure of exogenous and endogenous factors, inducingapproximately 104 defects per day (Slupphaug et al. 2003). Genetic information is transferred from onegeneration to the next one, therefore it’s principal to keep it intact. In the case of DNA damage, the cellmust imply reparation mechanisms (Sharova 2005). Unrepaired defects are fixed in the genome asmutations, or induce chromosome breaks. They can lead to defects at the level of cells, tissues, organs,degenerative changes, senescence and death of both unicellular and multicellular organisms (Sancar etal. 2004). The genomic integrity is protected by control points in G1 and G2 phase, which are able toslow the cell cycle until the elimination of the defect (Norbury, Hickson 2001).

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During the aging process, the accumulated mutations can result to functional defects, which canlead even to death (Sax 1962). Reparation mechanisms are able to repair the DNA damage duringontogenesis. The theory of mutation aging assumes an increasing number of unrepaired mutations inthe course of aging. Strehler (1962) and Ahlert (1971) however argue against this theory, proclaimingthat the higher frequency of mutations with age is a consequence, not the cause of aging.

The evidence about conserved genetic base of aging is an argument against this theory. The theoryof mutation accumulations assumes the selection pressure against genes which cause degenerativechanges in the old age being so weak, that they are accumulated in the genome. These genes shouldthen be random defects, accumulated in the genome, which contradicts the know facts about conservedgroups of genes connecrted with aging (Mitteldorf 2010). One such group encodes the components ofthe insulin/IGF signal path, which accelerates aging in the abundance of nourishment (Barbieri et al.2003). Inactivation of the CLK-1 gene in worms enhances the lifespan significantly (Johnson et al.1993), as well as the MCLK-1 gene in mice (Liu et al. 2005). Budovsky et al. (2007) elaborated the listof next conserved genes connected with aging.

1.4. Accumulation of mutations in plantsIn plants, the possibility of modular growth and replacing the old tissues with new structures, the

question of accumulation of mutations is most important in the meristemes. The meristemes presentalso the main difference between life strategies in plants. They divide many times during the life of theplant, and so they are exposed not only to the environmental mutagens, but to the possible mutagenousdefects during the mitosis as well. The meristematic cells of one-year plants undergo approximately 50 divisions in one life cycle, similarly to mammal cells (Otto, Walbot 1990). The meristematic cellsof perennial plants, however, undergo many more divisions during the life of the plant, increasing thepossibility of somatic mutations (Sax 1962).

The detection of mutations is possible with the use of pollen, meristematic tissues, saplings, andseeds. The use of pollen biological properties in the detection of the mutagenic potential of theenvironment is based on several basic assumptions. The first is the fact that the pollen grain is a haploidcell and lethal mutations originating in antecedent DNA replication express themselves by the changeof biological activity especially vitality of pollen. Pollen abortion can be caused by even small deletionsor point mutations. Most of the point mutations originating from the genotoxic impact of theenvironment are lethal. The genes with the mutations caused by environmental impact tend to berecessive form of heterozygous allele (Mulcahy, Johnson 1978). The other assumption is that in thecase the changes of pollen biochemistry were caused by physiological causes, all set of pollen grainsin anther would be affected (Murín, Mičieta 2000). For these studies, only diploid plants can be used.Mičieta and Murín (1997) studied the pollen abortivity of Pinus sylvestris and Pinus nigra asbioindicators for the environmental pollution, and found a significant increase of pollen abortivity inpolluted environment. Therefore, if mutations can induce aging, then the presence of mutagenes in theenvironment could also influence this process. Lanner and Connor (2001) studied the pollen viabilitydepending on age in Pinus longaeva Engelm., and found an age related trend, but the results were notstatistically significant. Bhattacharya (2005), however, found a statictically significant increase ofpollen abortivity with age in the species Anacardium excelsum L.

Thomas (2002) however claims that the aging of plants is not directly linked with the accumulationof genetic damage. Somatic mutations propagated through cell lines contribute to the higher frequencyof chimeras. Meristeme is an equivalent of a microbiological thermostat (Klekowski 1988) whereunfavourable mutations are suppressed through intraorganismal selection, while positive mutations canbe a source for increased adaptation ability (Gill et al. 1995, Orive 2001).

1.5. Aging of seedsSeeds are a special case of aging in plants, which could be put under the category of organ aging.

It can be assumed with near certainty, that this process is not programmed, as it would be against the

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life plan of the plant. Seed weight and viability can be measured as a mark of aging. Lanner and Connor(2001) observed no differenced in seed weight and viability depending on the age of the tree.

Other authors however, observed the aging of the seeds themselves. Inocenti and Avanzi (1971)assume, that the aging of seeds is caused by the accumulation of mutagenous metabolites. Floris andAnguilessi (1974) found a decreased activity of enzymes like catalase, peroxidase, cytochrome oxidaseand decarboxylase in older seeds, but also the loss of protein-synthetic capacity and increasedpermeability of membranes, leading to the reduction of sugar content, and content of other metabolicproducts. Ethanol and other automutagenes can be accumulated during the respiration. Murín andMièieta (2001) found a great individual variability in the aging seeds of V. faba. Aging significantlyinfluences germination and the root growth of the seedlings, as well as the chromosome damage. The frequency of chromosome aberrations was, however, lower than expected. These results suggest,that aging of seeds has greater impact in the physiological, than genetic level. In spite of theexpectation, the content of free endogenous cytokines zeatine and zeatine riboside has not decreased.The enzyme initiating the transformation of zeatine to dihydrozeatinecan play a key role in the vitalityreduction of old seeds. The indication of seed aging in V. faba is darkening of the testa. The seeds ofthe same age show individual variability, therefore it can be concluded that the aging process ischaracteristic for individuals, not for genetic lines or species.

Seeds of many plants are extremely tolerant to harsh environmental conditions provided they arein a state of dessication. In this dry state, their metabolic activity is drastically reduced to a very lowlevel (quiescence) while retaining their ability to germinate for considerable periods (Rajjou,Debeaujon 2008).

Murín and Mièieta (2001) also discovered that the aging of seeds depends on the storageconditions, especially from the relative humidity. Low water content in the seeds limits biochemicalprocesses, possible light reactions, and some oxidation processes. At the humidity of 30 % the effectof so-called ‘artificial aging’ occurs, and the seedlings of three years old seeds have the same rootlength as from ten years old, stored in normal conditions. This rate of humidity enhances the enzymaticreactions, delays the start of the first mitosis, but is not sufficient for the pre-replication excision repairof DNA damage. When the humidity is raised to 50 %, rejuvenation caused by the storage effect occurs.This humidity enables the reparation processes, and the storage prolongs the time available forreparation. The germination of 12 years old seeds after storage was the same as in 2 years old seeds(Murín, Mièieta 2001). Such treatments, which are based upon controlled hydratation of the seeds, arereferred to as “priming” During treatment seeds remain tolerant to desiccation because of incompletehydration and can be re-dried (Heydecker et al. 1973). Yet, paradoxically, a number of laboratorystudies conducted with dessication-tolerant (orthodox) seeds tend to show that low relative humidityand low temperature or cryopreservation might correspond to the optimal storage conditions to theoptimal storage conditions to improve seed life span (Villiers 1973; Rajjou, Debeaujon 2008).

There have been reported some extreme cases of seed preservation and longevity. The oldestdiscovered seeds determined by radiocarbon dating were the seeds of date (Phoenix dactylifera L.) –2000 years (Sallon et al. 2008), sacred lotus (Nelumbo nucifera) – 1300 years (Shen-Miller 2002), andcanna (Canna compacta) – 600 years (Lerman, Cigliano 1971).

2. Theories connected with the size of the organism

2.1. General theories connected with the size of the organismFinch (1990) introduced the term of “negligible aging” in the organisms which fulfil the following

assumptions: insignificant change in mortality rate with age, insignificant change in the reproductionrate after reaching reproductive maturity, insignificant degeneration of physiological functions withage, and insignificant decrease of resistance against diseases. Some examples of animal species withnegligible aging are: Greenland whale, giant tortoise, flatfish, sturgeon, lobster, etc. All of there

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organisms have one common trait – unlimited growth. Therefore the age can be deduced from the sizeof the organism, although there are some exceptions as well. The theory that growth limitation is themain cause of aging is relatively old (Minot 1908).

The high developed organisms are not able to maintain their physiological functions without a population of highly differentiated cells with no or very low proliferation activity (neurons,cardiomyocyts). In the same time, these cells face DNA damage that they are not able to repaircompletely. Cell division enables the replacement of damaged cells with a process similar to naturalselection at the cellular level. The experiments of Esipov et al. (2008) showed that contact inhibitionof proliferating cells induces changes in DNA, similar to those observable in aging organisms. Theseresults led to the opinion, that the development program itself is sufficient to explain aging (Khokhlov,2010). Therefore organisms without growth limitation are potentially immortal. The same applies tocell lines of transformed human and animal cells and germ lines of all organisms.

2.2. Theories connected with the size in plantsThe same assumption about growth limitation can be applied to all modular organisms including

plants. In trees, the relation between volume and height growth is not linear. For a maximum stability,the biomass would have to increase exponentially to maintain constant height growth (Ryan et al.2006). Therefore older trees slow their height growth and many of them assume a specific growth form– flattened top of the crown, thickening of primary branches, the branches of lesser order become moretwisted and gnarled (Ryan, Yoder 1997). There are numerous theories explaining the height growthlimitation, mostly based on the physical and hydraulic properties of trees: the hydraulic limitationhypothesis (Ryan, Yoder 1997), turgor limitation hypothesis (Koch et al. 2004), and sink limitationhypothesis (Ryan et al. 2006).

The trees of one species live up to a certain average age, but this age can substantially differ inindividuals of the species. The size and age that a tree can reach depends considerably on theenvironmental conditions. LaMarche (1969) assumes that the environmental conditions can haveinfluence on acquiring the specific growth form characteristic for old trees of the species Pinuslongaeva – restriction of cambial activity into a narrow part of the trunk circumference, which negatesthe size as a factor substituting physical age. Weathering and abrasion by soil particles graduallyremove the bark and cambium from the upper side of the roots, while the living part of the root remainson the bottom side. The same happens due to the abrasion by wind and snow in the aboveground partof the plant. The repeated loss of leaves and branches on the windward side can also suppress theactivity of cambium. Colder and dryer conditions also influence the resistance of the wood – theysuppress the pathogens, and lead to the production of more dense, highly lignified wood.

Also relatively short-lived trees can reach considerable age in specific conditions, as Larson (2001)shows on the example of Thuja occidentalis L. This species inhabits two kinds of habitats in NorthAmerica: wet swamp forests and stony cliffs, where it can withstand drought and extreme temperatures.In the wet forests it’s a short-lived pioneer tree, with the lifespan of 80, maximally 400 years. On theother hand, the individuals growing on the cliffs have a significantly different growth form andlifespan. In the comparison with the trees growing in favourable conditions, they are very small and,and have an asymmetric, gnarled trunk. Individuals with the trunk diameter under 40 cm, and age 1890years have been found. Collier and Boyer (1989) have refuted the theory of genetic difference of thesepopulations, so the differences are caused exclusively by environmental conditions.

Schulman (1958) formulated a hypothesis, that longevity can be induced by unfavourableconditions. However, Larner (2002) claims, that other long-lived tree species can reach an extreme agein favourable conditions. We can conclude that there are no specific environmental conditions leadingto longevity. For every tree species there are condition that enable it to reach the maximum age, butthey are specific for every species, based on the evolution history and life strategy.

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3. Theories based on the cell division

3.1. General theories of telomere shorteningSince the discovery of the function of telomeres, a new wave of aging theories based on a limited

number of cell divisions – Hayflick’s limit (Hayflick 1979) have been published. Telomeres are anessential part of all eukaryotic chromosomes as a protection against the activity of exonucleases andchromosome fusion. They are short, tandem repetitions, rich on guanine bases (Riha et al. 1998),shortened after every cell division due to the inability of DNA polymerases to completely replicate the5’ end of linear DNA with Okasaki fragments. However, in certain cells the length of telomeres ispreserved with the help of telomerase – a specific reverse transcriptase which uses its own subunit asa template (Blackburn 1991, Zakian 1995).

The consequence of telomere shortening accompanying the cell division is replicative senescence(Allsopp et al. 1995). Cells having the ability of unlimited division have a stabilized length of telomeresand high telomerase activity. These cells are practically immortal in vivo: germ line cells, some virus-transformed cells and 90 % of malign tumors (Kim et al. 1994). Organs of highly specialized animals,however, contain post-mitotic or slowly growing cells, and so the term “proliferation potential” doesn’thave sense for them. The cells capable of division almost never exhaust their potential during the lifeof the organism (Cristofalo et al. 1998). That leads to the conclusion, that telomere shortening is notthe primary cause of aging.

3.2. Apical meristemes and telomeres in plantsThomas (2002) assumes that apical meristemes are essential for the life span of plants. The

potential life span is unlimited, as far as the apical meristeme gives origin to new vegetative organs andsustains the cell line alternative to the stem cells in animals. The life span is determined with the timewhile the apical meristeme is able to produce new cells quicker than the proceeding wave of cell death(its form depends at the plant’s life form). In the case of trees and shrubs the dead tissues persist aswood. However, if a terminal set of organs is established together with the decrease in proliferationcapacity, the meristeme becomes determinated. The determination is usually associated with thetransformation of the organs from vegetative to generative. In the case of polycarpic plants at least onemeristeme at the end of the vegetation period remains non-determinated. Polycarpy versus monocarpyis, however, not only a qualitative characteristic. Many genes participating on the cell division, growth,senescence and maturation have been described. The life history is a result of quantitative relations inthe expression of these genes.

In most plants, telomeres contain of a repeating sequence of nucleotids (TTTAGGG)n (Ganal et al.1991). There are differences in the ontogenesis of plants and animals, which cause that the telomeresin plants keep their length (Riha et al. 1998). Plants don’t have a specialized line of germ cellsdifferentiated in early embryogenesis like animals. Every meristematic cell is potentially, throughseveral generations of cells, the precursor of a sex cell. Therefore every change in the nuclear genomeof the meristematic cell, including telomere shortening, can be carried over to sexual offspring.Telomere shortening would lead to the complete loss of telomeres after several generations.

Flanary and Kletetschka (2005) studied the length of telomeres and telomerase activity in Pinuslongaeva. They observed a cyclical pattern of telomere length with age, caused by the cyclic activityof telomerases, with the period approximately 1.500 years. The changes of telomere length were bestobservable in roots, while the pattern of telomerase activity was clearest in the needles. Activetelomerases have been found in all samples from different aged trees. This suggests that the meristemsof trees have unlimited replication capacity.

4. Theories of evolutionary compromises

These theories are based on the assumption, that the evolutionary case of aging is an unavoidablecompromise between preserving the soma, and other tasks necessary for fitness, like metabolism and

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reproduction. These theories have sense, only if these compromises are really necessary. On the otherhand, aging genes that offer no observable advantage in exchange have been discovered as well(Mitteldorf 2010).

4.1. Antagonistic pleiotropy The theory of antagonistic pleiotropy (Williams 1957) is a theory of the compromise between aging

and reproduction, based on the exchange of fitness in early life stages for the price paid in later stages.For many genes, their effect in old age is indeed balanced by their earlier positive effects, but thevalidity of this theory is question in the light of two facts: the positive influence of these genes is notuniversal, and the advantages that they offer are often just marginal for surviving and reproduction.This theory also strongly depends on the fact, if the compromises are necessary, and if longevity couldbe reached without sacrificing of other benefits. Experiments indicate that the compromises betweenlongevity and fertility are not a cause of existence of the aging genes, just its side effect (Mitteldorf2010). Lines of Drosophila with an increased longevity without a negative effect on other traits (witha positive effect on fitness, even) have been bred in laboratory (Leroi et al. 1994). Some of the aginggenes identified in Caenorhabditis elegans and other organisms have no other effect besides shorteningthe lifespan (Johnson et al. 1993).

4.2. Disposable soma theory This theory by Kirkwood (1977) is based on metabolic compromises. For the available energy,

there are competing metabolic demands – for reproduction, immunity defence etc. The somatic repairis only one of these demands, therefore the repair of damage can’t be perfect, and damage will beaccumulated, similar to the damage accumulation theory. Some experiments, however, speak againstthis theory: the mice which get less nourishment, so the energy available to them is lower, tend to havea longer lifespan then well-fed mice (Mitteldorf 2010). For many species, reproduction requires a significant energetic investment, so in the given conditions of available energy, it either takes placeor not. According to this theory, the survivorship curve would rise when the reproduction stops, andthen will drop further with the decrease of available energy from nourishment. However, the truth is,that the lifespan slightly increases through a wide scale of food deprivation. Several studies confirmthis positive correlation (Ricklefs, Cadena 2007).

4.3. Aging as deprivation syndrom Heininger (2002) promoted the view on aging as a universal pattern of reaction, programmed as

a prototypic answer of the organism on metabolic stress. It developed already in unicellular organismson the purpose of resistance or mutation adaptation to environmental stress. In the lack of resources thecells differentiate to spores, which are able to surpass unfavourable conditions, and the rest of the cellsperish. After the transition to multicellular organisms, the mechanism remain evolutionary fixed, withthe germ line of cells analogous to the surviving spores, and somatic cells programmed to perish afterthe end of reproduction. So aging developed in the world of limited resources, as an answer to the deathsentence proclaimed by the selection pressure and executed by the differentiation of germ cells.

4.4. Adaptative pleiotropy Adaptative pleiotropy is an alternate theory of evolutionary compromises, modified by Mitteldorf

(2010). It assumes that if we accept that aging itself is an evolutionary adaptation, it must be anadaptation on the group level, because it offers no advantages to the individuals. The question is whyindividual selection does not remove aging. A possible answer is, that the pleiotropy itself is anadaptation, selected as a tool to preserve the genetic basis for againg against the opposing force ofindividual selection. This theory is a possible explanation of the existence of antagonistic pleiotropy,not as reason of aging, but as a means of its preservation against short-term selection, which is blind tothe long-term advantages on the group level.

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4.5. Evolutionary compromises in plantsIn plants, the storage of resources has a great influence on the senescence (Thomas 2002). In the

case of many monocarpic plants, senescence is a result of exhaustion because of the nutrition demandsof seeds, but it is not a rule: suppressing the development doesn’t have to influence senescence, or caneven accelerate it. The male individuals of dioecious plants undergo terminal senescencesimultaneously with the female plants, which is an evidence of programmed character of this process.In sterile plants the evolution of delayed or accelerated senescence can be favoured depending on therelative importance of herbivory and competition in their life history.

5. Adaptative theories of aging

Multiple arguments against the above mentioned theories can support the view on aging asadaptation. Two forms of death are known in unicellular organisms – apoptosis and cell senescencethrough telomeres. These events are connected with stress response, and are advantageous for themicrobial community. Both forms are evolutionary conserved in higher organisms as well, as have beenalready stated in the theory of deprivation syndrom (Heininger 2002). Conserved groups of genesconnected with aging remain related also in distant taxa (Guarante, Kenyon 2000). Low additionalgenetic variance, which is a measure of variation in natural genotypes by the study of selection foraging, is also a mark of adaptation (Promislow et al. 1996).

Hormesis is a term used for a positive answer of an organism to low doses of stressor. It has beenproved, that the lack of food, just like other forms of stress, can lead to increased longevity (Luckey1999). This fact is a proof for the plasticity of lifespan under genetic control, and the possibility toprolong it without any compensation or side effects. The question is why this genetic program does nottake effect in favourable conditions (Mitteldorf 2010). Experiments with C. elegans and Drosophilashowed that besides the food availability, also its smell influencing the nerve system influences aging(Libert et al. 2007), similarly like obfuscating of the chemical sensor of C. elegans can lead toprolonged lifespan (Apfeld, Kenyon 1999).

Next argument for programmed aging as evolutionary adaptation is the programmed death ofsemelparous plants and animals after the end of reproduction. Many ecologists suppose this event is a form of adaptation at the group level (Crespi, Teo 2002). The idea of aging developing as anadaptation, to acceleration the adaptative changes in the population, has been rejected in the pastbecause of the long time necessary for such changes taking efect, observable only in evolutionary timescale (Williams 1966, Maynar Smith 1976). However, Gilpin (1975) proved that ecological interactionscan change the balance between selection for individual fitness and cooperation in the group.

5.1. Demographic theory of aging Mitteldorf (2006) presumes, that there is no intrinsic reason for the stability of ecosystems – to such

a measure, that demographic homeostasis must be an evolutionary adaptation. No species livesisolated, but interacts with the other species, and depends on them. Therefore ecological homeostasisbecomes a significant goal of the natural selection, competing with the tendency of maximalization theindividual reproduction outcome, and moderating individual fitness with its effect. This evolutionaryprocess is fast, because it influences the fitness itself, not the rate of its change as the theory about thesignificance of aging for the acceleration of adaptative changes in the population presumes. Senescencedirects the individual reproduction outcome in a way that leads to demographic homeostasis. Withoutthis direction, a tendency for rapid growth in favorable conditions, and collapse in unfavorableconditions would occur in the population. Aging moderates these effects by restriction of the lifespanof the individuals in a growing population, while it has no effect on declining populations where mostof mortality is caused by starvation. On the other side, hormesis increases the efficiency of senescencefor controlling the stability of population further, because it presses the mortality rate in theseconditions even lower. Another effect of population dynamics that can be important for the evolutionof aging is the morbidity and epidemics. Aging helps to regulate the population density and increase its

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diversity. A similar argument like the one about the influence of parasites and diseases on the evolutionof sexual reproduction according to the “hypothesis of the Red Queen” (van Valen 1973) is applicableto the evolution of aging. So the population dynamics can support the evolution of senescence in theopposition to individual selection.

5.2. Adaptative theories of aging in plantsPlant populations have certain specifics due to their modular nature. It is not entirely clear, what

can be considered a population, especially in the long-lived trees. Plants differ with the ability totransport the resources from one part of the plant to another. The rate of this ability is expressed bysectoriality – the rate of segregation of the transport paths. Its manifestation in trees can be observed inthe different coloring of individual branches. Mineral contain can differ up to 30 %. Hydraulic isolationof individual sectors creates differences in the traits inside the crown (Ellmore et al. 2006). Somaticmutations can also play a role in these differences. Ellmore at al. (2006) studied the rate of sectorialityin relation to the longevity of trees. They found significantly higher rates on sectoriality in the long-lived trees (36 times higher sectoriality in Quercus rubra than in Betula papyrifera). Larson (2001)measured hydraulic segmentation in the two different populations of Thuja occidentalis: the short-livedpopulation living in wet regions, and the long-lived population growing on dry cliffs. The slowlygrowing trees in unfavorable conditions have higher sectoriality than the population in favorableconditions. The practical effect is that the trees with high sectoriality can be viewed as a population of individual modules within the frame of one central axis (Larson 2001). The same is valid for Pinuslongaeva, and preliminary proves have been obtained for the species of gender Juniperus: J. virginianaL., J. communis L., J. oxycedrus L., J. phoenice L., and the gender Taxus, all with potential lifespan ofover 1.000 years. None of these species survives until high age with intact cambium. The highsectoriality enables the localization of potentially catastrophic events into one or several hydraulicsectors, while the rest remains intact, even grows more quickly, because the resources are dividedbetween a lesser numbers of living cells (Kelly et al. 1992).

Therefore the mechanism of aging in trees can be explained by demographic theory of agingapplied on an individual tree as a population of independent modules. The application of this theorydepends on the rate of sectoriality of the given species of woody plant. That makes trees remarkableorganisms, for which the processes of population dynamics can act with different impact within theframe of population as within the frame of an individual tree as well.

Discussion

Despite the number of published theories on aging in living organisms, still there remains muchunclear about this phenomenon. Most of the studies of aging focus on animal organisms, where theimplication to human aging is clearest. However, the oldest organisms in the world are plants. The oldest living trees are the bristlecone pines, Pinus longaeva growing in the White Mountains ineastern California and Snake Range in eastern Nevada. Schulman (1958) was the first to describe theirlongevity. The highest known age of a living tree is 4.843 years of a pine known under the name‘Methuselah’ in Schulman’s Grove in the White Mountains. It sprouted in the year 2832 BC. The highest counted number of tree-rings at all was found in a tree with the nickname ‘Prometheus’ onthe Wheeler Peak mountain in Snake Range, after it has been cut down for a dendrochronologic study– 4.862 in the height 2,5 m above ground level, so it is possible that the actual age of the tree was above5.000 years (Currey 1965). However, if we consider the possible clonality of trees, it gives us a newperspective of their age. The most famous clonal system is the growth of quaking aspen (Populustremuloides Michx.) in Wasatch Range, in the Rocky Mountains, Utah, named ‘Pando’ (‘I spread’ inLatin) (Grant 1993). It is a male clone of about 47.000 genetically identical trees which share a common root system, and cover the area of 43 ha. The age estimations differ from 10.000 years (sincethe last glacial) up to 1 million years (Mitton, Grant 1996).

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The aging of plants, and trees especially, is maybe even less studied and explained that aging ingeneral. Some trees live only a few decades, while others seem to be potentially immortal. There havebeen partial results in explaining the causes of longevity, growth limitation and aging on cellular level,but so far, no consistent theory of tree aging and longevity has been published. The present studiessuggest, that the mortality in trees is mostly caused by external influences, then internal aging, whichspeaks in the favour of the non-programmed theories of aging, although it doesn’t answer the questionif it is a universal principle or an exception. In monocarpic plants however, the senescence seems to beprogrammed, or at least controlled by the development of seeds.

An important characteristic of plants, that can bring further insight into the question of their agingand longevity, is their modularity and compartmentalisation. All plants are modular organisms with nogrowth limitation as in unitary organisms, although there are still other factors limiting their growth.The connection (or the lack it) between hydraulic paths within the plant can also lead to innersegmentation, as Ellmore et al. (2006) has shown. If we quantitate the rate of the segmentation from 0 to 1, we get a scale from total connection of hydraulic paths like in most herbaceous species, to thetotal segmentation, which could be identified with clonality. The range between these two extremes iscontinuous, which causes the different impact of negative exogenous causes on the surviving of thewhole plant. The closer to the end of the range, the lesser is the impact on the plant as a whole, sinceit remains isolated to a few almost independent sectors. Therefore the plants close to this end of thescale behave like a population of independent modules.

The extreme end of the scale – clonality – is a way to maintain undetermined growth, which is a condition for immortality according to Minot (1908) and Finch (1990), without the hydrauliclimitation of size. However, undetermined growth means also a great number of mitotic divisions,which increases the possibility of somatic mutations, causing the “mutation meltdown” of a clonalpopulation (Klekowski 2003). From the view of aging theories, there is one more importantcharacteristic of plants – they do not differentiate the line of germ cells. Therefore somatic mutationscan be carried over to the generative organs, and to gametes (Buss 1983). During clonal growth,mutations that influence sexual fitness negatively can be accumulated, while they have no influence onthe clonal growth itself (Eckert 2002, Klekowski 2003). Ally et al. (2003) found a significant decreaseof male fertility with age in the clonal systems of Populus tremuloides. It can be considered a form ofreproductive senescence, while the other characteristics typically connected with senescence remainunder selection pressure, and are not influenced by the age of the clonal system. This is evidence thata) somatic mutations occur in clonal systems, and b) natural selection acts as a force influencing theviability of the mutants. We can conclude that similar processes are take place in highly segmentedtrees, which behave as a population of independent clonal modules within one axis. The study ofLanner and Connor (2001) did not find significant differences between pollen viability of old andyoung individuals of Pinus longaeva, which means that the selection pressure for sexual fitness is stillpresent in this case. We can conclude that while clonality is a means of reproduction, and potentiallyunlimited, modularity within one axis is still specially limited by hydraulic characteristics. Thereforesexual reproduction must be preserved as a means of spreading the genetic material, and remains underselection pressure. Still somatic mutations occur, and can be influenced by intra-organismal selectionbetween modules, which can lead to possible chimerism within the plant.

The implication of these findings is that a single tree can behave as a population of modules, wherethe individual modules can undergo the processes of non-programmed aging, while the tree as a wholeremains almost unaffected. The situation is similar to animal population, where it is much easier topredict the maximum age of an animal, than the possible time of surviving for the whole population.This time is affected mostly by the conditions of the environment, and possible cathastrophic events.The same is valid for long-lived trees, where this very longevity can depend on the environmentalconditions, as it is in the case of Pinus lonageva (LaMarche 1969), or can be even caused by theseconditions as it is in the case of Thuja occidentalis, where they influence also the rate of hydraulicsegmentation (Larson 2001).

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Acknowledgements

The authors gratefully acknowledge financial support by VEGA (Project No. 1/0182/09).

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Abstrakt

Hlavnou otázkou teórií starnutia je, či je starnutie programované alebo nie a ako sa vyvinulo v evolúcii. Hoci tieto otázky neboli zatiaľ jasne zodpovedané, bolo publikovaných niekoľko skupínteórií, zaoberajúcich sa touto problematikou. Väčšina týchto teórií však neberie do úvahy rastlinnéorganizmy a ich špecifické vlastnosti, ktoré robia mechanizmy ich starnutia jedinečnými. Prvoutakouto vlastnosťou je klonalita a sektorialita, druhou je nediferencovanie zárodočnej línie buniek.Absencia zárodočnej línie je príčinou toho, že teloméry sa neskracujú a môže tiež viesť k prenosusomatických mutácií na sexuálne potomstvo. Naproti tomu sektorialita u stromov spôsobuje izoláciupotenciálne katastrofických udalostí do jednej časti stromu a robí ho populáciou viac-menej ne -závislých modulov v rámci jednej osi. Preto k procesom populačnej dynamiky, vrátane stárnutia, môžedochádzať pod rôznymi vpyvmi v rámci populácií i v rámci jednotlivých stromov.

Eva Brutovská, Karol Mičieta, Andrea Sámelová: Aplikácie teórií starnutia na rastlinné organiz -my s dôrazom na stromy

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PUBLISHED RECORDS OF THE GENUS CREPIDOTUS FROM SLOVAKIA

Soňa Jančovičová•

Comenius University in Bratislava, Faculty of Natural Sciences, Department of Botany, Révová 39, 811 02 Bratislava, Slovakia

Received 2 August 2011; Received in revised form 31 August; Accepted 5 October 2011

Abstract

A literature search was made to collect the publications in which the records of Crepidotus from Slovakia arementioned. In 60 publications of 40 authors, records of 16 species were published: Crepidotus applanataus, C. autochthonus, C. calolepis, C. carpaticus, C. caspari C. cesatii, C. crocophyllus, C. ehrendorferi, C. epibryus,C. kubickae, C. luteolus, C. macedonicus, C. mollis, C. subverrucisporus, C. variabilis and C. versutus. For eachspecies, the number of published records, the number of deposited specimens and the occurrence within thephytogeographical units of Slovakia are enumerated.

Key words: fungi, Agaricales, literature, excerption, herbaria

Introduction

Being interested in the genus Crepidotus, I have provided field observations (e.g. Ripková, Blanár2004, Ripková et al. 2007b), re-examined voucher specimens (Ripková 2002a), solved taxonomicpuzzles (Ripková et al. 2005, Ripková 2009) etc. Besides these, I also wanted to know what itspublication history in Slovakia is. Hence, I made a literature search to collect the publications in whichrecords of Crepidotus species are mentioned. My aim was to find out 1) the Crepidotus diversity inSlovakia, 2) the numbers of Crepidotus records from Slovakia, and 3) whether/where the voucherspecimens of the Crepidotus species are kept.

Material and Methods

The database created for the Checklist of Slovak fungi (Bacigálová, Lizoň 1999), later updated byAdamčík et al. (2003), and the book Mycoflora Slovaca by Škubla (2003) were used as bibliographicalsources. Several other works, mostly published during the years 2002–2010, were also excerpted. Thelists of taxa drawn up during some mycological forays and then deposited as a personal or institutionalmaterial, and theses, e.g. Msc thesis, were not accepted as reliable source of data in this work. Theexcerpted data were put into the “Database_Crepidotus_SJ” (i.e. authoress’s personal database; toolarge for publishing) in their original form (only minimal formal corrections were made).

For all records of Crepidotus species, the phytogeographical units of Slovakia were added ifmissing and unified according to Futák (1966). Southern and northern parts of Biele Karpaty Mts. unitwere not distinguished, as well Lúčanská Fatra and Krivánska Fatra of the Malá Fatra Mts. unit.

The nomenclature and taxonomic concept of Crepidotus species follow Senn-Irlet (1995), Pouzar(2005), Ripková (2009) and Ripková et al. 2005. In this study, there are only Crepidotus namesfollowing the recent (above mentioned) taxonomic concepts. The names/synonyms under which the

27

Acta Botanica Universitatis Comenianae Vol. 46, 2011

• Corresponding Author: Soňa Jančovičová; jancovicova@fns.uniba.sk

records were originally published can be traced in the “Database_Crepidotus_SJ”; e.g. C. caspari waspublished by Pilát (1954) as C. lundellii; C. kubickae by Kotlaba (1961) as C. sphaerosporus.

The herbaria abbreviations are cited in accordance with the Index Herbariorum (Holmgren et al. 1990).

Results

A literature search resulted in the selection of 60 works (published till the year 2010) in which therecords of the genus Crepidotus from Slovakia were mentioned. The publications were written by 40authors who collected data on 16 species: Crepidotus applanataus, C. autochthonus, C. calolepis, C. carpaticus, C. caspari C. cesatii, C. crocophyllus, C. ehrendorferi, C. epibryus, C. kubickae, C. luteolus, C. macedonicus, C. mollis, C. subverrucisporus, C. variabilis and C. versutus.

Undoubtedly, there is omitted literature that I have not excerpted (e.g. some difficult-to-tracearticles, some old and uninterpretable publications and/or simply forgotten ones). The numberspresented in the Tabs. 1 and 2 (the numbers of publications, records, specimens) are, therefore, onlyapproximative.

Based on the overview of the years of publications, Crepidotus records were published sporadicallytill the 80th of 20th century (1856, 1926, 1933, 1948, 1954), but more or less continually since then(1971, 1975–1978, 1982, 1984, 1987–1989, 1991, 1993, 1994–1997, 1999, 2002–2007, 2009, 2010).As it can be seen, the oldest record is from 1858, the newest one from 2010.

Concerning the publications in which Crepidotus records are mentioned, in the highest number ofthem (in more than 20 publications) C. applanatus, C. mollis and C. variabilis were published, in thelowest number (only in 1 or 2 publications) C. carpaticus, C. ehrendorferi and C. macedonicus (Tab. 1).

Summing up the number of records for each Crepidotus species, the highest number (more than100) is known for C. applanatus, C. mollis and C. variabilis; the lowest number (less than 5) for C. autochthonus and C. carpaticus (Tab. 2)

As to the phytogeographical units of Slovakia, the highest number of Crepidotus records (morethan 50) is known from the Podunajská nížina Lowland, Slovenské rudohorie Mts., Muránska planinaPlateau, Vihorlatské vrchy Mts. and Bukovské vrchy Mts.; the lowest number (only 1) from PohronskýInovec Mts., Slovenský raj Mts., Liptovská kotlina Basin and Šarišská vrchovina Mts.; no record ofCrepidotus species was hitherto published from Východoslovenská nížina Lowland, Považský InovecMts., Javorie Mts., stredné Pohornádie Mts., Chočské vrchy Mts., Belianske Tatry Mts., Javorníky Mts.and Čergov Mts. (Tab. 2).

While excerpting data, I also noted whether/where the voucher specimens were deposited. I havefound out that according to the published data, about 60 % of all Crepidotus records were documentedby specimens which are kept in the institutional herbaria BRA, BRNM, PRM, SLO, UPS as well as inthe private herbaria of S. Glejdura, S. Hagara, J. Herink, J. Hlaváček and J. Kuthan. Consequently, I verified the real existence of the specimens, however, only in the institutional herbaria of SLO, SAVand BRA, and private herbaria of L. Hagara and S. Glejdura; it resulted in high agreement (almost 95 %) (“Database_Crepidotus_SJ”). The numbers of specimens correspond to the numbers of publishedrecords, i.e. the highest number of specimens exist for C. applanatus, C. mollis and C. variabilis; the lowest for C. autochthonus and C. carpaticus.

I hope that the data I have excerpted and worked up in “Database_Crepidotus_SJ” will be useful toupdate the database of the checklist of Slovak fungi (Bacigálová, Lizoň 1999), the red list of Slovakfungi (Lizoň 2001), and, might be, for the new volume of Flora of Slovakia (cf. Paulech 1995,Bacigálová 2010). For this moment, my efforts are to continue adding the new Crepidotus records(published after 2010) as well as to revise the voucher specimens from Slovakia.

28

Tab. 1. The publications in which the records of the genus Crepidotus from Slovakia are mentionedcolumn “Crepidotus species” – for original names/synonyms, under which the records were published, the“Database_Crepidotus_SJ” (i.e. authoress’s personal database) is at disposalcolumn “No.” – the total number of publications

29

Crepidotus species Publications No.

C. applanatus Adamčík (1994), Adamčík et al. (2006), Aghová et al. (1988), Bolla (1856), Dermek (1976, 1977, 1978), Janitor (1994), Kopecká (1982), Kult (1991), Kuthan (1989), Kuthan et al. (1999), Lazebníček (1987), Lizoň, Kautmanová (2004), Pilát (1948), Ripková (2002a), Ripková, Blanár (2004), Ripková et al. 2007a, 2007b), Senn-Irlet (1995), Škubla (2003), Záhorovská (1997) 22

C. autochthonus Kuthan et al. (1999), Ripková (2002a), Škubla (2003) 3

C. calolepis Kotlaba et al. (1991), Kuthan et al. (1999), Pilát (1948), Ripková (2002a), Ripková, Blanár (2004) 5

C. carpaticus Škubla (1996) 1

C. caspari Adamčík et al. (2006), Blanár, Mihál (2002), Ďuriška (2010), Kotlaba et al. (1991), Kuthan et al. (1999), Pilát (1954), Ripková (2002a), Ripková, Blanár (2004), Ripková, Kučera (2002), Ripková et al. (2007b), Škubla (1995, 1996, 2003) 13

C. cesatii Adamčík et al. (2006), Blanár, Mihál (2002), Kuthan et al. (1999), Mihál et al. (2010), Pilát (1948), Ripková (2002a, 2009), Ripková, Blanár (2004), Ripková, Kučera (2002), Ripková et al. (2007a, 2007b), Škubla (2003) 12

C. crocophyllus Ďuriška (2010), Antonín et al. (1995), Ripková (2002a), Ripková et al. (2005, 2007b), Škubla (1994, 1995, 1997, 2003) 9

C. ehrendorferi Ripková, Glejdura (2010) 1

C. epibryus Adamčík, Hagara (2003a, 2003b), Ďuriška (2010), Kuthan et al. (1999), Ripková (2002), Ripková, Blanár (2004), Ripková et al. (2007b), Škubla (2003) 8

C. kubickae Glejdura, Kunca (2010), Kotlaba (1961), Ripková (2009) 3

C. luteolus Blanár, Mihál (2002), Kult (1989), Kuthan (1989), Kuthan et al. (1999), Lizoň (1977b), Ripková (2002), Ripková, Blanár (2004), Škubla (2003) 8

C. macedonicus Ripková (2002a, 2002b) 2

C. mollis Adamčík et al. (2006), Dermek (1977, 1978), Dermek, Michalko (1975), Fábry et al. (1975), Fellner (1994), Glejdura, Kunca (2010), Janitor (1994, 1997), Kotlaba (1961), Kult (1991), Kuthan (1989), Kuthan et al. (1999), Lazebníček (1987), Příhoda (1971), Pilát (1948), Ripková (2002a), Ripková, Blanár (2004), Ripková et al. (2007a, 2007b), Sak (1993), Záhorovská (1997), Záhorovská, Lišková (1996), Záhorovská et al. (1996), Škubla (2003), 25

C. subverrucisporus Adamčík et al. (2006), Ripková, Blanár (2004), Ripková et al. (2007b), Škubla (2003) 4

C. variabilis Adamčík et al. (2006), Aghová et al. (1988), Dermek (1977, 1978), Dermek, Michalko (1975), Janitor (1994, 1997), Kult (1991), Kuthan (1984), Kuthan et al. (1999), Lizoň (1977a, 1977b, 2006), Mihál (1997a, 1997b, 1997c, 1997d), Pilát (1926), Ripková (2002a), Ripková, Blanár (2004), Ripková, Kučera (2002), Ripková et al. (2007a, 2007b), Škubla (2003), Záhorovská (1997), Záhorovská, Jančovičová (1997), Záhorovská, Lišková (1996), Záhorovská et al. (1996) 28

C. versutus Adamčík et al. (2006), Pilát (1948), Ripková (2002a), Ripková, Blanár (2004), Ripková et al. (2007b) 5

30

Tab

. 2. T

he

nu

mb

er o

f p

ub

lish

ed/d

epos

ited

rec

ord

sof

th

e ge

nu

s C

repi

dotu

s in

th

e p

hyt

ogeo

grap

hic

al u

nit

s of

Slo

vak

ia

Bur

da M

ts.

Ipeľ

sko-

rim

avsk

á br

ázda

Fur

row

Slo

vens

ký k

ras

Kar

st

Záh

orsk

á ní

žina

Low

land

Dev

ínsk

a K

obyl

a M

ts.

Pod

unaj

ská

níži

na L

owla

nd

Koš

ická

kot

lina

Bas

in

Výc

hodo

slov

ensk

á ní

žina

Low

land

Bie

le K

arpa

ty M

ts.

Mal

é K

arpa

ty M

ts.

Pov

ažsk

ý In

ovec

Mts

.

Trí

beč

Mts

.

Str

ážov

ské

a S

úľov

ské

vrch

y M

ts.

Poh

rons

ký I

nove

c M

ts.

Vtá

čnik

Mts

.

Kre

mni

cké

vrch

y M

ts.

Poľ

ana

Mts

.

Šti

avni

cké

vrch

y M

ts.

Javo

rie

Mts

.

Slo

vens

ké r

udoh

orie

Mts

.

Mur

ánsk

a pl

anin

a P

late

au

Slo

vens

ký r

aj M

ts.

stre

dné

Poh

orná

die

Mts

.

Sla

nské

vrc

hy M

ts.

Vih

orla

tské

vrc

hy M

ts

Mal

á F

atra

Mts

.2/

0

C. applanatus

C. autochthonus

C. calolepis

C. carpaticus

C. caspari

C. cesatii

C. crocophyllus

C. ehrendorferi

C. epibryus

C. kubickae

C. luteolus

C. macedonicus

C. mollis

C. subverrucisporus

C. variabilis

C. versutus

Σ

3/3

6/3

1/0

1/1

2/2

4/2

1/0

1/0

4/3

6/6

16/1

5

1/0

33/3

0

2/0

1/1

1/0

1/1

4/4

6/6

8/6

3/3

3/3

6/4

1/1

1/1

3/1

4/4

21/2

0

11/1

1

1/1

8/7

1/1

1/1

1/1

2/2

2/1

15/1

4

9/9

3/3

8/4

1/1

28/2

6

1/1

1/1

8/8

5/5

3/3

1/0

1/1

11/9

4/4

6/6

1/1

1/1

3/3

2/0

3/1

5/5

1/1

3/3

1/1

1/1

1/1

1/1

8/8

2/2

2/0

7/5

4/2

12/9

3/3

2/1

8/6

2/0

2/1

1/1

1/1

21/1

5

11/1

0

16/1

3

1/0

8/1

6/4

2/0

3/3

4/4

5/4

4/4

8/5

1/1

7/3

5/3

3/1

14/1

4

11/5

5/2

2/1

10/3

3/1

9/0

25/2

5

1/1

4/4

1/0

6/6

1/1

3/3

15/1

2

14/1

1

34/2

1

18/1

3

54/4

3

3/3 -

25/2

3

28/1

5

- 9/3

3/2

1/1

2/1

2/2

15/1

2

49/3

5

-

70/6

2

85/7

6

1/0 -

18/1

95/9

0

6/2

31

Tab

. 2. T

he

con

tin

uat

ion

Veľ

ká F

atra

Mts

.

Cho

čské

vrc

hy M

ts.

Níz

ke T

atry

Mts

.

Záp

adné

Tat

ry M

ts.

Vys

oké

Tat

ry M

ts.

Bel

ians

ke T

atry

Mts

.

Tur

čian

ska

kotl

ina

Bas

in

Lip

tovs

ká k

otli

na B

asin

Spi

šské

kot

liny

Bas

ins

Javo

rník

y M

ts.

Záp

adné

Bes

kydy

Mts

.

Spi

šské

vrc

hy M

ts.

Šar

išsk

á vr

chov

ina

Mts

.

Čer

gov

Mts

.

Níz

ke B

esky

dy M

ts.

Buk

ovsk

é vr

chy

Mts

.

Σ

C. applanatus

C. autochthonus

C. calolepis

C. carpaticus

C. caspari

C. cesatii

C. crocophyllus

C. ehrendorferi

C. epibryus

C. kubickae

C. luteolus

C. macedonicus

C. mollis

C. subverrucisporus

C. variabilis

C. versutus

Σ

4/1

1/0

1/1

1/1

2/1

1/1

1/1

18/1

7

110/

88

2/2

4/3

1/1

20/1

8

1/1

1/1

2/1

55/4

9

1/0

3/0

47/4

0

1/0

2/2

42/3

58/

8

2/1

7/1

40/3

05/

5

1/0

3/1

15/8

15/1

5

2/0

1/0

1/0

2/0

1/0

1/1

3/3

2/0

1/1

2/2

10/2

137/

83

1/0

17/1

5

2/0

1/0

2/2

6/2

114/

68

3/3

16/1

5

11/2 - 2/0

2/1

4/0 - 4/3

1/0

3/2 - 4/4

2/0

1/1 - 4/3

58/3

3

646/

480

Acknowledgements

I am grateful to Pavel Lizoň and Viktor Kučera (Institute of Botany SAS, Bratislava) for help withliterature. Drahoš Blanár (Administration of the National Park of Muránska planina, Revúca) is acknowledged for identification of some phytogeographical units of Slovakia. Thanks also go to JanHolec (National Museum, Mycological Department, Praha) and Marica Zaliberová (Institute of BotanySAS, Bratislava) for reading and commenting on the manuscript. The Slovak Grant Agency VEGA(grants no. 2/0062/10, 2/0028/11) supported this study.

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Kuthan, J., 1989: Soupis makromycetů sbíraných na průzkumných exkurzích (s myk. NDR) ve dnech 3. IX. – 9. IX. 1988 a na exkurzích „Setkání českých a slovenských mykologů pod Tatrami“ dne 10. IX. – 18. IX. 1988 na lokalitách v oblastiVysokých Tater a Nízkych Tater a v Slovenském rudohoří. In: Kuthan J. (ed.), Houby rašelinišť a bažinatých lesů v Československu, p. 63-90, ČVSM pri ČSAV, Praha.

Kuthan, J., Adamčík, S., Antonín, V., Terray, J., 1999: Huby Národného parku Poloniny. Správa národných parkov SR,Správa Národného praku Poloniny, Liptovský Mikuláš, Snina.

Lazebníček, J., 1987: Huby. In: Vološčuk I., Terray J. (eds.), Chránená krajinná oblasť Vihorlat, p. 56-61, Príroda, Bratislava.Lizoň, P., 1977a: Exkurzie 1. mykologických dní na Slovensku. Správy hubárskej poradne, 4: 2-8.Lizoň, P., 1977b: VI. celoštátna mykologická konferencia v Pezinku. Správy hubárskej poradne, 5: 2-5. Lizoň, P., 2001: Červený zoznam húb Slovenska. 3. verzia. In: Baláž D., Marhold k., Urban P. (eds.), Červený zoznam rastlín

a živočíchov Slovenska, Ochr. Prír. 20 (Suppl.): 6-13.Lizoň, P., 2006: Macrofungi collected during the 9th mycological foray in Slovakia. Catathelasma, 7: 17-33.Lizoň, P., Kautmanová, I., 2004: Fungi collected during the 21st European Cortinarius foray. Catathelasma, 5: 23-35.Mihál, I., 1997a: Mykoflóra nezmiešanej bučiny v blízkosti hlinikárne v Žiari nad Hronom. Spravodajca slovenských

mykológov, 16: 24-25. Mihál, I., 1997b: Mykoflóra nezmiešanej bučiny v oblasti s miernym imisným vplyvom. Spravodajca slovenských

mykológov, 17: 21-22. Mihál, I., 1997c: Zoznam makromycétov zistených v okolí ipeľského predmostia. Spravodajca slovenských mykológov, 17:

22-23.Mihál, I., 1997d: K poznaniu mykoflóry lužných lesov a xerotermných dubín v okolí Ipľa. In: Urban P., Hrivnák R. (eds.),

Poiplie, p. 7-10. Slovenská agentúra životného prostredia, Banská Bystrica.Mihál, I., Glejdura, S., Blanár, D., 2010: Makromycéty (Zygomycota, Ascomycota, Basidiomycota) v masíve Kohúta

(Stolické vrchy). Reussia, 6: 1-44.Paulech, C., 1995: Erysiphales (Múčnatkotvaré). Flóra Slovenska, X/1: 1-292.Pilát, A., 1926: Les Agaricales et Aphyllophorales des Carpathes Centrales. Bull. Soc. Mycol. France, 42: 81-120.Pilát, A., 1948: Evropské druhy trepkovitek. Crepidotus Fr. Atlas hub evropských, 6: 1-77.Pilát, A., 1954: Houby na Polaně u Detvy na Slovensku. Čas. Nár. Mus., Odd. Přír., 123: 156-163.Příhoda, A., 1971: Choroby dřevin v Tatrách v letech 1966–1967. In: Marček A., Strnka M. (eds.), Zborník prác o Tatran -

skom národnom parku 13, p. 383-395, Osveta, Martin.Pouzar, Z., 2005: Klíč na určování našich trepkovitek (Crepidotus) a poznámky k nim. Mykol. Listy, 93: 1-9. Ripková, S., 2002a: Genus of Crepidotus in the herbarium of the Slovak National Museum − Natural History Museum,

Bratislava. Acta Rer. Natur. Mus. Nat. Slov., 48: 27-39. Ripková, S., 2002b: Crepidotus macedonicus, a new species for central Europe. Mycotaxon, 84: 111-118.Ripková, S., 2009: Crepidotus kubickae – a forgotten name. Mycotaxon, 110: 271-281.Ripková, S., Blanár, D., 2004: Výskyt druhov rodu Crepidotus na Muránskej planine a v priľahlej oblasti Slovenského

rudohoria. Reussia 1, Supplement, 1: 49-67.Ripková, S., Adamčík, S., Kučera, V., 2007a: Contribution to the knowledge of macrofungi in the Cerová vrchovina Mts.,

Juhoslovenská kotlina Basin and the Laborecká vrchovina Mts. Acta Botanica Universitatis Comenianae, 43: 15-23.Ripková, S., Adamčík, S., Kučera, V., Palko, L., 2007b: Fungi of the Protected Landscape Area of Vihorlat / Huby

Chránenej krajinnej oblasti Vihorlat. Institute of Botany of the Slovak Academy of Sciences / Botanický ústav Slovenskejakadémie vied, Bratislava.

Ripková, S., Aime, M. C., Lizoň, P., 2005: Crepidotus crocophyllus includes C. nephrodes. Mycotaxon, 91: 397-403.Ripková, S., Glejdura, S., 2010: Crepidotus ehrendorferi in Slovakia and taxonomic notes on related species. Czech Mycol.,

61: 175-185.Ripková, S., Kučera, V., 2002: Mykofloristický kurz. Spravodajca Slovenskej mykologickej spoločnosti, 27: 17-19.Sak, V., 1933: Pět neděl na houbách na Slovensku II. Časopis československých houbařů, 13: 57-58.Senn-Irlet, B., 1995: The genus Crepidotus (Fr.) Staude in Europe. Persoonia, 16: 1-80.Škubla, P., 1994: Vzácnejšie nálezy roku 1993. Spravodajca slovenských mykológov, 3: 18-19.Škubla, P., 1995: Vzácnejšie nálezy roku 1994. Spravodajca slovenských mykológov, 7: 25-27.Škubla, P., 1996: Vzácnejšie nálezy roku 1995 (pokračovanie). Spravodajca slovenských mykológov, 12: 26-28Škubla, P., 1997: Vzácnejšie nálezy roku 1996. Spravodajca slovenských mykológov, 16: 27-29.Škubla, P., 2003: Mycoflora Slovaca. Vlastným nákladom, Šaľa.Záhorovská, E., 1997: Huby (Fungi). In: Feráková V., Kocianová E. (eds.), Flóra, geológia a paleontológia Devínskej

Kobyly, p. 58-68, Litera s. r. o. pre APOP, Bratislava.

33

Záhorovská, E., Jančovičová, S., 1997: Mykoflóra alúvia rieky Rudavy. Spravodajca slovenských mykológov, 15: 15-19. Záhorovská, E., Lišková, D., 1996: Mykolóra Devínskej Kobyly medzi obcami Devín a Devínska N. Ves. Spravodajca

slovenských mykológov, 11: 20-22.Záhorovská, E., Lišková, D., Vozárová, M., 1996: Mykoflóra ostrova Sihoť a Slovanského ostrova. Spravodajca

slovenských mykológov, 14: 21-23.

Abstrakt

Predmetom práce bolo spracovanie literatúry, v ktorej sa uvádzajú nálezy rodu Crepidotus zoSlovenska. V 60 publikáciách od 40 autorov boli publikované nálezy 16 druhov: Crepidotusapplanataus, C. autochthonus, C. calolepis, C. carpaticus, C. caspari C. cesatii, C. crocophyllus,C. ehrendorferi, C. epibryus, C. kubickae, C. luteolus, C. macedonicus, C. mollis, C. subverrucisporus,C. variabilis and C. versutus. Pre každý druh je zistený počet publikovaných nálezov, početherbárových položiek a jeho výskyt v rámci fytogeografických celkov Slovenska.

Soňa Jančovičová: Publikované nálezy rodu Crepidotus zo Slovenska

34

BRYOPHYTES OF MEDIEVAL RUINS IN THE PROTECTED LANDSCAPE AREA MALÉ KARPATY MTS.

Daniela Uhereková Šmelková1•, Katarína Mišíková1, Anna Kubinská2

1Comennius University in Bratislava, Faculty of Natural Sciences, Department of Botany, Révová 39, 811 02 Bratislava, Slovakia

2Botanical Institute of Slovak Academy of Science, Dúbravská cesta 9, 845 23 Bratislava 4, Slovakia

Received 8 August 2011; Received in revised form 24 August; Accepted 4 October 2011

Abstract

Presented study brings a short overview of bryophytes of ten medieval ruins in the Protected Landscape Area MaléKarpaty Mts. Overall, 46 species (4 liverworts and 42 mosses) were recorded. The most species rich was orderHypnales. Comparing with other Slovak castle ruins, in the Malé Karpaty area the bryophyte diversity is significantly higher; however, the cover number is distinctly lower. The most interesting finding is that of Rhynchostegium rotundifolium, included in the Red list of Bryophytes of Slovakia (EN) and the EuropeanBryophyte Red Book (R).

Key words: castle ruins, Bryophyta, Slovakia, epilithic

Introduction

Old castles and their ruins form specific habitat for many species of bryophytes, which are mostlypioneer organism on stones and walls. Bryophyte diversity on these sites is affected by several factors:atmospheric elements, substrate type, degree of shadow, moisture and anthropogenic influence, e.g. airpollution, an important factor on ruins in urban environment (García-Rowe, Saiz-Jimenez 2002).

Within Europe, comprehensive research on bryophytes of castle ruins have not been realized up tonow, except in Slovakia, where a basic bryological research of these sites is performed at present(Uhereková Šmelková, Mišíková 2010).

Medieval ruins in the Malé Karpaty Mts. were not studied in past. On the other hand, some data arefrom the localities near by the ruins Pajštún (e.g. Bäumler 1884, Baumgartner 1901, Peciar 1981) andOstrý Kameň (Peciar 1990). Important information brings also report of Baumgartner (1901) from theruin Biely Kameň including the occurrence of rare moss Rhynchostegium rotundifolium; the presenceon this locality was confirmed by our study.

Material and methods

Field research was realized from October 2010 to June 2011. It was focused on epilithicbryophytes, samples were collected from castle walls, bricks and castle stones. The study includes eightcastle ruins, one monastery ruin and one ruin of a guard tower (Tab. 1). Frequency is calculated byformula: Fr (%) = (n1/N) • 100, (n1 is the number of sites where the species was recorded; N is the totalsites number).

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Acta Botanica Universitatis Comenianae Vol. 46, 2011

• Corresponding Author: Daniela Uhereková Šmelková; daniela.smelkova@gmail.com

The names of taxa and taxonomic hierarchy follow the works by Söderström et al. (2007) and Hillet al. (2006). Threat categories were established according to Kubinská et al. (2001) and ECCB (1995).

Herbarium specimens are deposited in the SLO herbarium (Comennius University in Bratislava,Faculty of Natural Sciences, Herbarium of the Department of Botany).

Geological structure of the Malé Karpaty Mts. consists of pre-alpine basis, alpine-age and mesozoicnappes (Plašienka et al. 1991), prevailing substrates are granites, basalts and limestones. Substrate ofobserved sites is mostly alkaline, predominatly by calcareous stones, brickwork and plaster.

Tab. 1. Index of observed sitesExplanations: Name/type: ca – castle, ga – guard tower, mo – monastery; Stage: A – reconstruction/conservation, B – basicpreservation works, C – desolate

Results and discussion

Overall, 46 epilithic bryophyte species from ten Carpathians castle ruins (Tab. 2) were recorded. Ondamp and wet sites, there was observed the highest bryophyte diversity. However, most of the evaluatedsites showed xerothermic character with the high number of xerophilic species. It seems that shadedegree probably does not appear as an important factor for bryophyte diversity.

Tab. 2. The occurrence and frequency of bryophytes on observed sitesExplanation: for observed sites see Tab. 1

36

No.

1

2

3

4

5

6

7

8

9

10

Name/type

Biely Kameň/ca

Branč/ca

Devín/ca

Dobrá Voda/ca

Dračí Hrádok/ga

Katarínka/mo

Korlátka/ca

Ostrý Kameň/ca

Pajštún/ca

Plavecký hrad/ca

Village/city

Svätý Jur

Podbranč

Bratislava

Dobrá Voda

Borinka

Dechtice

Cerová

Buková

Borinka

PlaveckéPodhradie

GPS coordinates

N 48° 15’ 66, 20” E 17° 11’ 75, 40”

N 48° 44’ 02, 80” E 17° 28’ 06, 00”

N 48° 10’ 50, 30” E 16° 58’ 71, 80”

N 48° 36’ 34, 13”E 17° 31’ 42, 78”

N 48° 15’ 14, 40”E 17° 06’ 40, 68”

N 48° 33’ 17, 30”E 17° 32’ 09, 33”

N 48° 34’ 31, 61”E 17° 23’ 08, 99”

N 48° 31’ 19, 70”E 17° 22’ 18, 65”

N 48° 16’ 53, 00”E 17° 40’ 97, 40”

N 48° 29’ 37, 55”E 17°16’ 07, 36”

Alt.(m a.s. l.)

290

475

212

320

370

245

455

588

486

424

Date oforigin

1270–1295

1297

864

1263

13th century

1646

1278

13th century

13th century

1270

Date of destruction

1663 destroyed Osmans

end of 17th century –burned down

1809 destroyed byNapoleons army

1762 burned down

destroyed in 15th centuryby Hussites

1787 forcibly desertedafter convent nullification

desolated in 18th century

17th century destroyedduring Rakozi revolt

1810 destroyed byNapoleons army

desolated in 18th century

Areasize(m2)

4200

7500

67000

8400

100

2500

3600

4500

6800

5175

Stage

B

A

A

C

C

A

B

C

B

B

species observed sites frequency (%)

Abietinella abietina 6, 9 20Amblystegium subtile 1 10Amblystegium serpens 1, 6, 9 30Anomodon viticulosus 1, 4, 7, 8, 9 50Barbula unguiculata 7 10Brachythecium albicans 2 10Brachythecium rutabulum 3, 4, 7, 8, 10 50Brachythecium salebrosum 10 10

Tab. 2. Continuation

The most frequent species were Tortula muralis (Fr = 100%), Homalothecium sericeum (Fr = 90%),Anomodon viticulosus (Fr = 50%), Bryum capillare (Fr = 60%), Hypnum cupressiforme (Fr = 70%) andEncalypta vulgaris (Fr = 60%). Many of them (e.g. Bryum argenteum, B. capillare, Ceratodonpurpureus, Didymodon fallax, Grimmia pulvinata, Homalothecium sericeum, Orthotrichum anomalum,Syntrichia ruralis, Tortula muralis) are typical as for walls as for historical sites; they were found e.g. inthe archaeological site Tilmen Höyük (Ezer et al. 2008). Within the orders, the most species rich wasHypnales (43 %), the lowest frequency have species from orders Jungermanniales (1 %), Metzgeriales(1 %), Dicranales (1 %) and Porellales (1 %) (Fig. 1).

The highest species diversity was observed on the site Biely Kameň (43 species), the lowestdiversity on the sites Katarínka (17) and Dračí Hrádok (19) (Fig. 2). As the ruins of the monasteryKatarínka are under the complete reconstructions work and the former guard tower Dračí Hrádok is indesolate constitution, the low number of species diversity is not surprising. It is notable that the area of

37

species observed sites frequency (%)

Brachytheciastrum velutinum 1, 2, 5, 9 40Bryoerythrophyllum recurvirostrum 3, 6, 8, 10 40Bryum argenteum 2, 3, 8, 10 40Bryum caespiticium 2 10Bryum capillare 1, 2, 3, 8, 9, 10 60Ceratodon purpureus 2, 3, 9, 10 40Didymodon fallax 3, 7 20Didymodon rigidulus 2, 3, 4 30Encalypta streptocarpa 2, 5, 7, 10 40Encalypta vulgaris 2, 3, 5, 8, 9, 10 60Grimmia pulvinata 2, 3, 7 30Homalothecium lutescens 2, 4, 5, 7, 9, 10 60Homalothecium philippeanum 1, 2, 4, 5 40Homalothecium sericeum 1, 2, 3, 4, 6, 7, 8, 9, 10 90Hypnum cupressiforme 1, 2, 4, 6, 8, 9, 10 70Hypnum vaucheri 5 10Isothecium alopecuroides 4 10Leucodon sciuroides 6 10Metzgeria furcata 1, 4, 10 30Mnium marginatum 4 10Mnium stellare 1, 5, 8 30Neckera besseri 8 10Neckera complanata 1, 4 20Orthotrichum anomalum 1, 2, 4, 8, 10 50Orthotrichum diaphanum 2 10Plagiochila porelloides 4, 8 20Plagiomnium cuspidatum 1 10Plagiomnium rostratum 8 10Pohlia nutans 3 10Porella platyphylla 1 10Pseudoleskeella catenulata 4, 6 20Pseudoleskeella nervosa 1, 4 20Rhynchostegium rotundifolium 1 10Schistidium apocarpum 1, 2, 7, 10 40Syntrichia ruralis 2, 7, 8, 9, 10 50Thuidium tamariscinum 9 10Tortula muralis 1, 2, 3, 4, 5, 6, 7, 8, 9, 10 100Tortula subulata 1, 8 20

latter one site is only 100 m2, which is almost twenty times lower then others ruins. In case of the siteBiely Kameň there are probably several factors influencing the species diversity, such as forest coveron almost all of the site, no recent reconstruction works and low tourist activities. The confirmation ofan endangered moss Rhynchostegium rotundifolium (EN) on the site Biely Kameň (Baumgartner 1901)is important for current knowledge on distribution of rare species in Slovakia.

Fig. 1. Percentage of species in particular orders

Fig. 2. Species diversity on observed sites

Comparing with bryophyte diversity of other studied Slovak castle ruins (Uhereková Šmelková,Mišíková 2010), the species diversity in the Malé Karpaty is higher. In average 33 species wererecorded on each site, in comparison to 13 species on previously evaluated castle ruins. The reason forthat can be the absence of more detailed observation. On the other hand, the cover number on thosesites is distinctly higher. Bryophytes that have not been recorded on other Slovak castles till now areAmblystegium subtile, Bryum caespiticium, Homalothecium philippeanum, Mnium marginatum,Orthotrichum diaphanum, Plagiomnium rostratum, Pseudoleskeella nervosa and Rhynchostegiumrotundifolium.

38

Hyp

nales

Pott

iale

s

Bry

ales

Enca

lypta

les

Grim

mia

les

Orthotr

ichal

esDic

ranal

es

Junger

man

niale

s

Met

zger

iale

s

Pore

llale

s

50

45

40

35

30

25

20

15

10

5

0

Bie

ly K

ameň

Bra

nčO

strý

Kam

eňPla

veck

ý hra

dDobrá

Voda

Paj

štún

Dev

ínKorl

átka

Dra

čí H

rádok

Kat

arín

ka

45

40

35

30

25

20

15

10

5

0

Ten observed medieval objects of Protected Landscape Area Malé Karpaty Mts. represent a different spectrum of ruin types. They differ from each other in the view of area size and stage (threeof them are desolated, three are conserved and four are under basic preservation works). Most of thesecastle ruins are conserved just thanks to many volunteers that repairing them during holiday. However,there are still many objects in dilapidated stage. Research of bryophytes on castle ruins is in early stageof elaboration and deserves more scientific attention in future.

Acknowledgements

We thank Miroslav Mišík (Medical University, Vienna) for fruitful discussion on draft manuscriptand review of the final text.

References

Baumgartner, J., 1901: Bryologische Exkursionen in das Gebiet der Pressburger Karpathen. Verh. Vereins Natur-Heilk.,Pressburg, 13: 17-23.

Bäumler, J., 1884: Die Moosflora von Pressburg in Ungarn. Oesterr. Bot. Z. 34: 46-49, 96-99. ECCB, 1995: Red Data Book of European Bryophytes. European Committee for Conservation of Bryophytes, Trondheim.Ezer T., Kara R., Çakan H., Düzenli A., 2008: Bryophytes on the archeological site of Tilmen Höyük, Gaziantep (Turkey).

Int. J. Bot. 4, 3: 297-302.García-Rowe, J., Saiz-Jimenez, C., 2002: Lichens and bryophytes as agents of deterioration of building materials in Spanish

cathedrals. Int. Biodeterior. 28, 1-4: 151-163.Hill, M. O., Bell, N., Bruggeman-Nannenga, M. A., Brugue´ S. M., Cano, M. J., Enroth, J., Flatberg, K. I., Frahm, J.-

-P., Gallego, M. T., Garilleti, Guerra, J., Hedenas, L., Holyoak, D. T., Hyvönen, J., Ignatov, M. S., R., Lara, F.,Mazimpaka, V., Munõz, J., Söderström, L., 2006: An annotated checklist of the mosses of Europe and Macaronesia.J. Bryol. 28: 198-267.

Kubinská, A., Janovicová K., Šoltés R., 2001: Červený zoznam machorastov Slovenska. In: Baláž, D., Marhold, K., Urban,P. (eds.), Červený zoznam rastlín a živočíchov Slovenska. Ochr. Prír. 20 (Suppl.): 31-43.

Peciar, V., 1981: Bryoflóra ohrozených oblastí Slovenska I. Acta Fac. Rerum Nat. Univ. Comenianae, Format. Protect. 6:105-118.

Peciar, V., 1990: Fytogeograficky významné zjavy v bryoflóre Malých Karpát. Biológia, Bratislava, 45, 9: 693-697.Plašienka , D., Michalík, J., Kováč, M., Gross, P., Putiš, M., 1991: Paleotectonic evolution of the Malé Karpaty Mts.:

An overview. Geol. Carpath. 42, 4: 195-208.Söderström, L., Urmi, E., Váňa, J., 2002: Distribution of Hepaticae and Anthocerotae in Europe and Macaronesia.

Lindbergia 27: 3-47.Uhereková Šmelková D., Mišíková K., 2010: Stručný prehľad machorastov vybraných hradov a zrúcanín na Slovensku.

Bryonora 46: 51-55.

Abstrakt

Nasledujúci príspevok prináša stručný prehľad bryoflóry desiatich stredovekých zrúcanín v oblastiCHKO Malé Karpaty. Zaznamenali sme 46 druhov machorastov (4 pečeňovky a 42 machov). Druhovonajbohatší bol rad Hypnales. V porovnaní s ostatnými zrúcaninami slovenských hradov, diverzitamachorastov na území Malých Karpát je výrazne vyššia ako na iných slovenských zrúcaninách hradov,kde je však vyššia celková pokryvnosť. Najzaujímavejší je nález druhu Rhynchostegium rotundifolium,ktorý je zaradený v Červenom zozname machorastov Slovenska (EN) a v Červenej knihe machorastovEurópy (R).

Daniela Uhereková Šmelková, Katarína Mišíková: Machorasty na stredovekých ruinách v oblasti Chránenej krajinnej oblasti Malé Karpaty

39

SOME SOIL CYANOBACTERIA AND ALGAE OF THE ZÁHORSKÁ NÍŽINA LOWLAND(SOUTHWEST SLOVAKIA)

Zuzana Drongová•, Ľubomír Kováčik

Comenius University in Bratislava, Faculty of Natural Sciences, Department of Botany, Révová 39, SK–81102 Bratislava, Slovakia

Received 16 September 2011; Received in revised form 6 October; Accepted 7 October 2011

Abstract

The soil phycoflora from the Záhorská nížina lowland was studied at 4 sampling sites – two sites on the grasslandsoil, one on the forest sandy soil and one on the open sandy site, where the cyanobacteria and algae in conjunctionwith mosses and fungi compose biological soil crusts which aid to stabilize the topsoil cover and protect soil fromwater and eolian erosion. Altogether 21 taxons of cyanobacteria and algae were identified in the locality. The mostfrequent species in the sampling sites were from division Chlorophyta (13) and less from division Stramenopila(3), Charophyta (3) and Cyanobacteria (2). During whole study the linear drawings and photos of found specieswere made.

Key words: soil cyanobacteria, soil algae, biological soil crust, sandy soil.

Introduction

Terrestrial cyanobacteria and algae are microscopic phototrophic organisms living in the firstseveral centimetres of soil. Remarkable is the ability of cyanobacteria to fix atmospheric nitrogen,change it to acceptable form for higher plants and increase its content in the soil (Bardgett 2005).Cyanobacteria together with algae take part in formation of biological soil crusts, connecting soilparticles together and protect soil against eolian and water erosion (Fletcher, Martin 1948, Belnap et al.2003). Hereby they help to maintain moisture in the higher soil layers and therefore improve conditionsfor seed germination and growth of vascular plants (Brotherson, Rushforth 1983, Ashley, Rushforth1984).

The objective of this study was to investigate dominant species of soil cyanobacteria and algaefrom locality Sekule – Mláky II at the Záhorská nížina lowland and preparation of biological materialto subsequent research of hydrological soil properties.

Material and methods

Location and sampling dataThe investigated area is located near the village of Sekule, the Záhorská nížina lowland, SW

Slovakia (48° 37’ 10” N, 17° 00’ 07” E) at elevation of 152 m a. s. l. Four locations were chosen forsampling (Fig. 1).

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Acta Botanica Universitatis Comenianae Vol. 46, 2011

• Corresponding Author: Zuzana Drongová; drongova@fns.uniba.sk

Fig. 1. Location map: (a) permanent green area, (b) permanent green area without surface vegetation, (c) forest site,(d) open sandy site

The first sampling site, SEK-1a, is a permanent green area with abound non-forest psamophyticvegetation (Fig. 2a). The most frequent species were from the family Poaceae (Agrostis capilaris andCynodon dactylon) and than Achillea millefolium, Acetosella vulgaris, Anthemis ruthenica,Convolvulus arvensis, Lepidium ruderale, Plantago lanceolata and Potentilla sp. (Lichner et al. 2005).The second sampling site, SEK-1b, is a permanent green area without surface vegetation (the plantcover was removed to the two-centimetre depth of the soil) (Fig. 2b). The third sampling site, SEK-2,is the forest sandy soil under Pinus sylvestris grove (Fig. 2c). The fourth sampling site, SEK-3, is theopen sandy site (Fig. 2d) and the both sampling sites, the third and the fourth, are partly covered by thelichens (Cladonia sp. div.), mosses (Dicranum polysetum, Ditrichum heteromallum, Hypnumcupressiforme, Polytrichastrum fumosum, Polytrichum piliferum) and occasionally grasses(Corynephorus canescens) (Šomšák et al. 2004). The soil microscopic fungi on these sites (Alternariaalternate, Aspergillus fischeri, A. galucus, A. niger, Humicola fuscoatra, Penicillium aspergilloides, P. decumbens, Trichoderma viride, T. kongii (Lichner et al. 2007, Kalivodová et al. 2008) whichtogether with cyanobacteria and algae form biological soil crusts on the top layer of the sandy soil andhelp to protect it from eolian and water erosion.

The climate of this area is continental with an average annual temperature is between 9 and 10 °C(Šťastný et. al 2002a), in summer tends to consist of long hot and dry spells interspersed with intenserainfall (Faško et al. 2008), average annual rainfall rate is 550 mm (Faško, Šťastný 2002). During thehottest month (July) the temperature is between 20 and 21 °C (Šťastný et al. 2002b), during the coldestmonth (January) the average month temperature does not fall under 2 °C (Šťastný et al. 2002c). Thesoil at sampling sites has sandy and sandy loam texture (Šály, Šurina 2002) classified as aeolian sandor regosol formed on aeolian sand (MKSPS, 2000). More detailed physical and chemical properties ofsoil from the site in Sekule Mláky II are given in the study Lichner et al. 2005.

42

Fig. 2. Sampling sites: (a) permanent green area, (b) permanent green area without surface vegetation, (c) forest sitecovered with needles, (d) open sandy site partly covered with mosses and biological soil crusts

Laboratory methodsSoil samples from investigated locality were taken into plastic sample tube in August 2010 and

March 2011. The soil samples from each sampling site were handled in laboratory dual way: a) The soil samples were spread onto agar plates with cultivation medium Z after Zehnder (Staub

1961) in Petri dishes. b) The soil samples were diluted in liquid Z medium in glass tubes and incubated at temperature

25 °C, under the irradiation of 48.8 µmol m-2 s-1 provided by five Narva Warm White lamps LT18W/530-030 for 14 days. Afterwards samples were inoculated onto agar plates in Petri dishes withcultivation medium Z in Petri dishes.

In both cases cyanobacteria and algae from Petri dishes were isolated into the new cultivationmedium and multiply isolated in order to obtain pure cultures of cyanobacteria and algae. If the diatomswere found, organic matter was cauterized in 30% H2O2 and mounted into the Pleurax to obtainpermanent slides.

On the other site, the samples were taken in monthly intervals from March to July 2011 from coverslips which were spreading on the top layer of the soil. After a month, exposed face of each taken coverslips (the part turned to the soil) was brushed on the agar plates in Petri dishes. Along with theorientation prospecting of microscopical growth on cover slips through light microscope were made.

43

All Petri dishes were exposed at temperature 25 °C, under the irradiation of 48.8 µmol m-2 s-1. After 10to 14 days cyanobacteria and algae were isolated under a stereomicroscope. Separate colonies werechosen and transferred on the slant agar in tubes, Erlenmeyer flasks or on agar plate into the Petri dishesand later re-isolated to obtain unispecific cultures of cyanobacteria and algae.

Fig. 3. The line drawings of found phycoflora: (a) Leptolyngbya sp., (b) Phormidium autumnale, (c) Klebsormidiumsubtile, (d) Xanthonema montanum, (e) Tribonema minus, (f) Stichococcus bacillaris, (g) Choricystis minor, (h)Mychonastes zofingiensis, (i) Eustigmatos polyphem

During whole study the linear drawings of found species were made (Fig. 3). All isolates were studiedunder optical microscope Leica DM 2500 using differential interference contrast (DIC), a Leica DFC 290HD camera and LAS 3.5.0 software were employed to document the observed objects (Fig. 4).

Species determination was made using standard references, mainly Ettl, Gärtner (1995), Lokhorst(1996), Andreyeva (1998), Belnap, Lange (2003). All strains are stored at the working culturecollection of cyanobacteria and algae of the Laboratory of experimental phycology, Department ofBotany, Faculty of Natural Sciences, Comenius University in Bratislava.

Results

During the observed periods following taxonomic groups in the locality were found with numbersof taxons in the brackets: Cyanobacteria (2), Chlorophyceae (4), Trebouxiophyceae (9),Klebsormidiophyceae (3), Xanthophyceae (2), Eustigmatophyceae (1).

As indicated this summary, the main taxons of algae in the sampling sites were from divisionChlorophyta: Bracteacoccus minor (Chodat) Petrova, Chlorella sp., Chloroidium elipsoideum(Gerneck) Darienko et al., Choricystis minor (Skuja) Fott, Diplosphaera cf. chodatii Bialosuknia,Geminella sp., Mychonastes zofingiensis (Dönz) Kalina et Punčochářová, Podohedra bicaudataGeitler, Radiosphaera minuta Herndon, Scotiellopsis oocystiformis (J.W.G. Lund) Punčochářová et Kalina, Stichococcus bacillaris Nägeli, S. undulatus Vinatzer and Tetracystis sp.

44

a b c d

e

10 µm

f

g

h

i

Fig. 4. Some of found cyanobacteria and alga species: (a) Leptolyngbya sp., (b) Phormidium autumnale, (c) Stichococcus bacillaris, (d) Scotiellopsis oocystiformis, (e) Klebsormidium subtile, (f) Tribonema minus, (g) Eustigmatos polyphem, (h) Radiosphaera minuta

45

From division Stramenopila there were found species: Eustigmatos polyphem (Pitschmann)Hibberd, Tribonema minus (Wille) Hazen, Xanthonema montanum (Vischer) Silva, and a fewrepresentatives of Bacillariophyceae in sampling site SEK-1a which were not determined in detail in this study.

From division Charophyta there were found species: Klebsormidium dissectum (Gay) Ettl et Gärtner, K. subtile (Kützing) Tracanna et Tell, K. mucosum (Boye Petersen) Lokhorst in Lokhorst etW. Star.

From division Cyanophyta there were found representative of genera Phormidium Kützing ex Gomont, and Leptolyngbya Anagnostidis et Komárek.

This sandy area is still in progress and various types of ecological succession (from primarysuccession on the bare soil) are observed here. The samplings were realized during autumn and springup to early summer and we assume that the number of found species will be growing within a year aswell as during onward succession.

The abundance was also influenced by the moisture of the studied site. According to Kalina, Váňa(2010) moisture is main limiting factor for terrestrial species and their live cycles often depends onslight horizontal rainfall.

All observed taxa are typical terrestrial species with wide range of appearance in the soils oftemperate climatic area. Isolated cultures will be used for next study and special laboratory experimentsaimed to study hydrophysical characteristics of the soil of this locality.

Tab. 1. List of species recorded in selected sampling sites in the Záhorskánížina Lowland

Acknowledgement

We would like to thank Daniela Uhereková Šmelková (Comenius University in Bratislava,Department of Botany) for help with determination of mosses in sampling site SEK-3. The research

46

Taxon / Sampling siter

Leptolyngbya sp.

Phormidium autumnaleBracteacoccus minorChlorella sp.

Chloroidium elipsoideumChoricystis minorDiplosphaera cf. chodatiiEustigmatos polyphemGeminella sp.

Klebsormidium dissectumK. mucosumK. subtileMychonastes zofingiensisPodohedra bicaudataRadiosphaera minutaScotiellopsis oocystiformisStichococcus bacillarisS. undulatusTetracystis sp.

Tribonema minusXanthonema montanum

SEK-1a

+

+

+

+

+

+

+

+

+

+

-

+

+

+

+

-

+

+

-

+

+

SEK -1b

+

+

+

+

-

+

+

+

-

+

-

+

+

-

-

-

+

-

-

+

-

SEK-2

-

-

+

+

-

+

-

-

-

+

-

-

+

-

-

-

+

-

-

-

-

SEK-3

+

-

+

+

+

+

+

+

+

+

+

+

+

-

+

+

+

+

+

-

-

was supported by the Scientific Grants Agency VEGA as part of project No. 2/0073/11 and byComenius University Grand UK/146/2011. The authors thank to a reviewer for his valuable commentsto the manuscript.

References

Andreyeva, V. M., 1998: Terrestrial and aerophilic green algae (Chlorophyta: Tetrasporales, Chlorococcales,Chlorosarcinales). NAUKA. St. Peterburg.

Ashley, J., Rushforth, R. S., 1984: Growth of soil algae on top soil and processed oil shale from the Uintah Basin, Utah,U.S.A. Reclamation and Revegetation Research, 3: 49-63.

Bardgett R., 2005: The Biology of Soil. A community and ecosystem approach. University of Lancaster, Lancaster, UK.Belnap, J., Lange, O. L. (eds.), 2003: Biological Soil Crusts: Structure, Function, and Management. Springer Publisher,

Berlin.Brotherson, J. D., Rushforth, S. R., 1983: Influence of cryptogamic crusts on moisture relationships of soil in Navajo

National Monumrnt, Arizona. Great Basis Naturalist, 43: 73-78. Ettl, H., Gärtner, G., 1995: Syllabus der Boden-, Luft- und Flechtenalgen. Gustav Fischer Verlag, Stuttgart.Faško, P., Šťastný, P., 2002: Priemerné ročné úhrny zrážok [Average annual rainfall] 1 : 2 000 000. Atlas krajiny Slovenskej

republiky. Ministerstvo životného prostredia SR, Bratislava, Slovenská agentúra životného prostredia, Banská Bystrica.Faško, P., Lapin, M., Pecho, J., 2008: 20-year extraordinary climatic period in Slovakia. Meteorol. Čas., 11: 99-105.Fletcher, J. E., Martin, W. P., 1948: Some effects of agae and molds in the rain-crust of desert soils. Ecology, 29: 95-100.Kalina, T., Váňa, J., 2010: Sinice, řasy, houby, mechorosty a podobné organismy v současné biologii. [Cyanobacteria, algae,

fungi, mosses and other organisms in present biology.] Karolinum, Praha.Kalivodová, E. (ed.), 2008: Flóra a fauna viatych pieskov Slovenska. [Flora and fauna of drift sands in Slovakia.] VEDA,

Bratislava.Lichner, Ľ., Nižnanská, Z., Faško, P., Šír, M., Tesař, M., 2005: Vplyv rastlinného pokryvu a počasia na pôdnohydrologické

parametre vodoodpudivej pôdy v lokalite Mláky II pri Sekuliach [The impact of plant cover and weather on soil-hydrological parameters of a water repellent soil at the locality Mláky II at Sekule]. Acta hydrol. Slovaca, 6: 321-329.

Lichner, Ľ., Hallett, P. D., Feeney, D., Dugová, O., Šír, M., Tesař, M., 2007: Field measurement of the impact ofhydrophobicity on soil water transport under different vegetation over time. Biologia, 62: 537-541.

Lokhorst, G. M., 1996: Comparative Taxonomic Studies on the Genus Klebsormidium (Cyanophyceae) in Europe. GustavFischer Verlag. Stuttgart.

MKSPS (2000): Morfogenetický klasifikačný systém pôd Slovenska. Bazálna referenčná taxonómia. [Morphogeneticclassification system of soils in Slovakia. The basal reference taxonomy] VÚPOP, Bratislava.

Šály, R., Šurina, B., 2002: Pôdy [Soils] 1 : 500 000. Atlas krajiny Slovenskej republiky. Ministerstvo životného prostrediaSR, Bratislava, Slovenská agentúra životného prostredia, Banská Bystrica.

Šomšák, L., Šimonovič, V., Kollár, J., 2004: Phytocoenoses of pine forests in the central part of the Záhorská nížinaLowland. Biologia, 59: 101-113.

Šťastný, P., Nieplová, E., Melo, M., 2002a: Priemerná ročná teplota vzduchu [Average annual air temperature] 1 : 2 000 000.Atlas krajiny Slovenskej republiky. Ministerstvo životného prostredia SR, Bratislava, Slovenská agentúra životnéhoprostredia, Banská Bystrica.

Šťastný, P., Nieplová, E., Melo, M., 2002b: Priemerná teplota vzduchu v júli [The average temperature in July] 1 : 2 000 000.Atlas krajiny Slovenskej republiky. Ministerstvo životného prostredia SR, Bratislava, Slovenská agentúra životnéhoprostredia, Banská Bystrica.

Šťastný, P., Nieplová, E., Melo, M., 2002c: Priemerná teplota vzduchu v januári [The average temperature in January] 1 : 2 000 000. Atlas krajiny Slovenskej republiky. Ministerstvo životného prostredia SR, Bratislava, Slovenská agentúraživotného prostredia, Banská Bystrica.

Staub, R., 1961: Ernährungsphysiologisch-autökologische Untersuchungen an der planktischen Blaualge Oscillatoriarubescens DC. Schweiz. Z. Hydrol., 23: 82-198a.

Abstrakt

Pôdna fykoflóra Záhorskej nížiny sa študovala na štyroch odberových miestach: dve miestapredstavovali na zatrávnenú pôdu, jedno na lesnej piesočnatej pôde a jedno na otvorenej piesočnatejploche, kde cyanobaktérie a riasy v spojení s machmi a hubami tvoria biologické pôdne krusty, ktorénapomáhajú stabilizovať povrch pôdy a chrániť ho pred vodnou a veternou eróziou. Spolu bolo

47

v lokalite identifikovaných 21 druhov cyanobaktérií a rias, najviac zastúpené druhy na odberovýchmiestach pochádzali z oddelenia Chlorophyta (13) a menej z oddelení Stramenopila (3), Charophyta (3)and Cyanobacteria (2). Počas celého štúdia boli zhotovované líniové kresby a fotografie zistenýchdruhov.

Zuzana Drongová, Ľubomír Kováčik: Niekoľko pôdnych cyanobaktérií a rias Záhorskej nížinyna juhozápade Slovenska

48

FILAMENTOUS GREEN ALGA KLEBSORMIDIUM FLACCIDUMAS A LABORATORY MODEL FOR THE STUDY OF DORMANCY

Jana Segečová1•, Josef Elster2, Ľubomír Kováčik1

1Comenius University in Bratislava, Faculty of Natural Sciences, Department of Botany, Révová 39, 811 02 Bratislava, Slovakia

2Academy of Sciences of the Czech Republic, Institute of Botany, 379 82 Třeboň; University of South Bohemia, Faculty of Science, Branišovská 31, 370 05 České Budějovice, Czech Republic

Received 14 September 2011; Received in revised form 5 October; Accepted 7 October 2011

Abstract

Filamentous green alga Klebsormidium flaccidum (Kütz.) P. C. Silva, Mattox et W. H. Blackw. (Klebsormidiales,Streptophyta), strain Kaštovská 2002/45, isolated from cryoconite sediment in Svalbard (Arctic, Ny-Ålesund,Austre Brøggerbreen) was used for experimental study of dormancy as a survival strategy. In unfavourableconditions, this alga is able to form morphological dormant stages, and thereby retain its viability underenvironmental stress. Slow air desiccation was used as a stressor.

Key words: Klebsormidium, green algae, dormancy

Introduction

Species of the genus Klebsormidium represent one of the most widespread groups of aero-terrestrialgreen algae in the world, with distribution ranging from polar to tropical habitats (Karsten et al. 2010,Nagao et al. 2008). Its taxonomy is still a challenge because of its morphological simplicity and probablehigh-hidden cryptic diversity. Many ultrastructural and molecular studies confirm the position ofKlebsormidium in the streptophycean lineage along with higher plants, and often highlight its highecological importance in aeroterrestrial habitats (Karsten et al. 2010). The representatives of the greenalga Klebsormidium have been well studied taxonomically by Lokhorst (1996) and he confirmed thatKlebsormidium belongs to the charophycean algae. In addition, Klebsormidium is an adequate laboratorymodel for experimental studies because of its easy cultivation in laboratory conditions. Dormancy is anadaptation to survive unfavourable conditions with the word “dormant” being a synonym for inactive.The most remarkable characteristic of the dormant stage is the significantly reduced or even undetectablerate of metabolism (Mulyukin et al. 1997). Changes in the rates of physiological processes are combinedwith additional protective mechanisms against various stresses. These include changes in membrane andcell envelope structure, changes in membrane and cell fluidity and the production of various specialprotective substances (Elster 1999). Dormancy can also be defined as a survival strategy, where cellshave a higher resistance to different stressful conditions. Finally, dormant cells must retain viability afterretun into favourable conditions (Mulyukin et al. 1997).

Various Klebsormidium strains from polar regions exhibit desiccation tolerance (Elster et al. 2008).The aim of this study is to reveal how Klebsormidium cells form their dormant stages.

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Acta Botanica Universitatis Comenianae Vol. 46, 2011

• Corresponding author: Jana Segečová; jana.segecova@gmail.com

Material and methods

The experimental alga Klebsormidium flaccidum (Kütz.) P. C. Silva, Mattox et W. H. Blackw.,strain Kaštovská 2002/45 isolated from Svalbard (Arctic, Ny-Ålesund, Austre Brøggerbreen,cryoconite sediment) and deposited in the Algal Culture Collection Institute of Botany Academy of Sciences of the Czech Republic in Třeboň (www.butbn.cas.cz/ccala/index.php) was used for thedesiccation experiment.

The cultures of this strain were grown in Petri dishes on agar plates with medium Z according toZehnder in Staub (1961) in an air-conditioned cultivation room at temperature ca. 20 °C. Illuminationwas provided by cool fluorescent lamps (48.6 μmol. m-2.s-1) with 12:12 h light-dark cycles.

The morphological characteristics of the strain phenotype, the cell wall structure and thickness,cytoplasm texture, pyrenoid morphology, starch grains and other storage material components werestudied under the JENAMED II (Carl Zeiss Jena) optical microscope. Cell types able to developdormant forms were especially searched for. Linear drawings and microphotographs using the LeicaDM 2500 optical microscope with Nomarski contrast, a Leica DFC 290 HD camera and LAS 3.5.0software were employed to document the observed objects.

Water was evaporated through a cotton plug from an experimental Erlenmeyer flask for ninemonths, so that a visible mark of approximately three quarters of agarized Z medium original volumewas apparent. The Klebsormidium flaccidum culture gradually exhausted available nutrients from theagarized medium and agar and this studied culture desiccated. The strains were then re-inoculated ontofresh agar plates with a Z medium and exposed to the same culture conditions as at the commencementof the experiment.

During this long term desiccation, we visually checked the colour of the algal biomass, andmicroscopically monitored the morphological characteristics of cells in the filaments and finallyprovided drawings and microphotographs.

Results

At the beginning of the experiment, the fresh growing culture on agar indicated the “best-fit”condition. Figures 1A, and 2A illustrate the cytomorphology of the cells and filaments of theKlebsormidium flaccidum fresh culture. Cells containing visible parietal chloroplasts and evidentpyrenoids clearly showed these vital characteristics.

After nine months of drying and starvation, the algal growth exhibited a yellow-green visualappearance. Figures 1B and 2B illustrate the character of filaments showing changes in theirmorphology; here, the equally formed or slightly bent filaments became tortuous with numerous bendsand bulges. The cell wall thickened, and there was a significant reduction in chloroplast to a shrunkenformation in the middle of the cell.

After re-inoculation of this alga onto fresh agar plates, the first hint of revitalization of growth wasvisibly noticeable in 3–5 days. This gradually increased and within the next few days the growth wasin the same optimal condition as at the commencement of the experiment (Figs 1C and 2C). Thisexperiment certainly confirmed the potency of the rapid revitalization ability of desiccatedKlebsormidium filamentous alga.

Discussion

The ability of Klebsormidium to withstand starvation and desiccation was described even in veryearly studies. Mattox (1971) described relatively unspecialized cells in Klebsormidium flaccidum thatremained viable filaments after air drying for more than 2 years. Morison & Sheath (1985) showed thatafter periods of desiccation, filaments of Klebsormidium sp. changed into unusual cell types; H-shaped

50

thick cell walls with refractive granules inside cytoplasma. The cell structure of Klebsormidiumrivulare, a drought-tolerant species, changed after exposure to drought; carbohydrate and lipid contentsincreased, and in contrary, protein content decreased. In the aquatic Klebsormidium rivulare, akinetesproduction was experimentally stimulated by desiccation. Thick, rigid cell walls have been describedin desiccated cells, indicating that this species increases cell wall thickness to withstand desiccation.

Fig. 1. Modification of the character of the Klebsormidium flaccidum filaments by desiccation andformation of dormant stage, A – filaments at the beginning of experiment, B – filaments after 9 monthsof desiccation, C – filaments after revitalization

Fig. 2. Microphotographs illustrate the modification in character of the Klebsormidium flaccidum filaments bydesiccation and formation of the dormant stage, A – filaments at the beginning of experiment, B – filaments after 9months of desiccation, C – filaments after revitalization. Scale bar 20 µm

In Klebsormidium flaccidum the accumulation of various low-molecular-weight carbohydrates andamino acids, together with ultrastructural changes such as chloroplast enlargements and a reduction invacuole size, have been described (Nagao et al. 2008). Ultrastructural changes of cold-acclimated cellsare also associated with the development of freezing-tolerance in Klebsormidium flaccidum. The vacuole size clearly decreased and the cytoplasmic volume increased in cold-acclimated cells. The accumulation of soluble sugars and amino acids in the enlarged cytoplasm of cold-acclimated cellscould efficiently protect many cell components. A recent study by Nagao et al. (2008) showed that thephysiological basis for cold acclimation in Klebsormidium flaccidum is represented by theaccumulation of amino acids and polysaccharides. Ultrastructural changes were examined in cold-acclimated cells, where deposits of oil droplets and an alteration of the pyrenoids with accumulation ofstarch grains were demonstrated.

51

A B C

A B C

In contrast to the mechanical deformation of Klebsormidium crenulatum desiccated cells, most ofthe cell organelles remain visibly intact, although altered (Holzinger et al. 2011). An accumulation oflarger irregular electron-dense compartments in the cytoplasm and the chloroplast is typical, togetherwith an increased number of plasto-globules within the chloroplast. Green algae cultured underunfavourable conditions often exhibit an accumulation of lipids in the cytoplasm. One of the moststriking observations from this study is the deformation of the cross walls between individual cells. In the SEM and TEM preparations, they appeared quite flexible. This allowed shrinkage of cell widthsto about 60% of their initial size, while the cell lengths were not affected by this desiccation.

In addition, Tribonema bombycinum, a freshwater alga from Tribophyceae, was reported to havesimilar features in the production of its dormant stages (Nagao et al. 1999). It transformed fromvegetative cells into akinetes during starvation in prolonged aquaculture in diluted medium, or inculture medium with particular nutrient deficiency. During starvation the vegetative cells, which had alarge central vacuole in the protoplasm and thin cell walls, finally changed to akinetes. These akinetescontain small vacuoles and oil droplets in the protoplasm and thick cell walls. However, akiniteformation was not facilitated in T. bombycinum akinetes, when desiccation was effected at the lowtemperature of 4 °C.

Acknowledgements

The study was supported by VEGA grant No. 1/0868/11, by UK grant No. 149/2011 and by TheGrant Ministry of Education of the Czech Republic INGO LA 341 and Kontakt ME 934.The authorsthank the reviewer for comments on the early version of this manuscript.

References

Elster, J., 1999: Algal versatility in various extreme environments. In: Seckbach, J., (ed.), Enigmatic microorganisms and lifein extreme environments, p. 215-227, Kluwer Academic Publishers, Dordrecht, The Netherlands.

Elster, J., Degma, P., Kováčik, Ľ., Valentová, L., Šramková, K., Batista Pereira, A., 2008: Freezing and desiccation injuryresistance in the filamentous green alga Klebsormidium from the Antarctic, Arctic and Slovakia. Biologia, 63: 843-851.

Holzinger, A., Lütz, C., Karsten, U., 2011: Desiccation stress cause structural and ultrastructural alterations in theaeroterrestrial green alga Klebsormidium crenulatum (Klebsormidiophyceae, Streptophyta) isolated from an alpine soilcrust. J. Phycol., 47: 591-602.

Karsten, U., Lütz, C., Holzinger, A., 2010: Ecophysiological performance of the aeroterrestrial green alga Klebsormidiumcrenulatum (Klebsormidiophyceae, Streptophyta) isolated from an alpine soil crust with an emphasis on desiccation stress.J. Phycol., 46: 1187-1197.

Lokhorst, G. M., 1996: Comparative taxonomic studies on the genus Klebsormidium (Charophyceae) in Europe. Crypt.Studies, 5: 1-132

Mattox, K. R., 1971: Zoosporogenesis and resistant-cell formation in Hormidium flaccidum. In: Parker, B. C., Brown, R. M.Jr. (eds.), Contributions in Phycology, p. 137-144, Allen Press, Lawrence, Kansas.

Morison, M. O., Sheath, R. G., 1985: Responses to desiccation stress by Klebsormidium rivulare (Ulotrichales, Chlorophyta)from a Rhode Island stream. Phycologia, 24: 129-145.

Mulyukin, A. L., Lusta, K. A., Gryaznova, M. N., Babushenko, E. S., Kozlova, A. N., Duzha, M. V., Mityushina, L. A.,Duda, V. I., El’-Registan, G. I., 1997: Formation of resting cells in microbial suspensions undergoing autolysis.Microbiology, 66: 32-38.

Nagao, M., Arakawa, K., Takezawa, D., Yoshida, S., Fujikawa, S., 1999: Akinete formation in Tribonema bombycinumDerbes et Solier (Xanthophyceae) in Relation to Freezing Tolerance. J. Plant Res., 112 : 163-174.

Nagao, M., Matsui, K., Uemura, M., 2008: Klebsormidium flaccidum, a charophyceaen green alga exhibits cold acclimationthat is closely associated with compatible solute accumulation and ultrastructural changes. Plant, Cell and Environment,31: 872-885.

Staub, R., 1961: Ernährungsphysiologisch-autökologische Untersuchungen an der planktischen Blaualge Oscillatoriarubescens DC. Schweiz. Z. Hydrol., 23: 82–198a.

52

Abstrakt

Vláknitá zelená riasa Klebsormidium flaccidum (Kütz.) P. C. Silva, Mattox et W. H. Blackw.(Klebsormidiales, Streptophyta), kmeň Kaštovská 2002/45 izolovaný z kryokonitového sedimentu zoSvalbardu (Arktída, Ny-Ålesund, Austre Brøggerbreen) sa použil na experimentálne štúdiumdormancie ako stratégie prežitia.V nepriaznivých podmienkach je táto riasa schopná tvoriť morfo -logické dormantné štádiá a takto si zachovať vitalitu aj počas pôsobenia environmentálneho stresu. Akostresor sa použilo pomalé vysušovanie na vzduchu.

Jana Segečová, Josef Elster, Ľubomír Kováčik: Vláknitá zelená riasa Klebsormidium flaccidumako laboratórny model na štúdium dormancie

53

MONTANE SYCAMORE BEECH FOREST IN THE OSOBITÁ MASSIF (WESTERN TATRAS) – PHYTOCOENOLOGICAL AND ECOLOGICAL ACCOUNT

Danica Černušáková•

Comenius University in Bratislava, Faculty of Natural Sciences, Department of Botany, Révová 39, 811 02 Bratislava, Slovakia

Received 23 March 2011; Received in revised form 10 September; Accepted 5 October 2011

Abstract

Montane sycamore beech forests occur in the Osobitá massif at the forest timber line (800–1300 m). We report theresults of phytocoenological and ecological analysis of these communities which belong to the association Cortuso-Fagetum. In this paper, the association is documented by 11 relevés, total species are 115 and average species are41. Three ecological factors are important for the prosperity of these forests: high air humidity, medium acidity(pH) and moisture of soil. These forests are sparse, open stands and of low growth, but they are fairly resistantagainst acid rain and strong wind. They do not form continuous stands, but rather limited islands.

Key words: beech, sycamore, montane forest, phytocoenology, ecological account, Osobitá massif

Introduction

Scientists, foresters and environmentalists in the High Tatra Mts. are trying to solve the challengeposed by the wind disaster from 2004. One of the still debated questions is whether to restore, cultivateand revitalize the woods using standard forestry techniques or to leave the process to the nature, i.e.,without any human interference. One of the possible routes leading to answers can be careful study ofthose natural forest communities which are least affected by human activity. Forest stands from Osobitámassif (Orava region in northern Slovakia) are an example of such community. These montane beechstands are especially interesting model systems because they span the range from 800 m up to timberline around 1300 m and often occupy habitats experiencing extreme climatic and orographicconditions. They do not exist as continuous stands, rather forming islands that are probably rest ofprevious communities. The forest communities of association Cortuso-Fagetum occur in the mountainlevel of Osobitá around the timber line in the altitudes 800–1300 m. The aim of this paper is to givesome indications on state of health of mixed forests from Osobitá based on the long-term study.

Communities of montane sycamore beech forests from subaliance Aceri-Fagenion were described bymany authors from different mountains of Europe: Klika (1927) reported from limestone of the Veľká FatraMts. the association Fagetum carpaticum (calcicolum) cortusae, later Klika (1936) modified it as theassociation Fagetum montanum carpaticum cortusae and in 1949 as the Abieto-Fagetum carpaticumcortusae. Hadač (1969) described similar communities from limestone of Belianske Tatry as theassociation Acero-Fagetum. Fajmonová (1982a, b) described montane sycamore forest growing onlimestone of Westcarpatian of Slovakia as the association Cortuso-Fagetum. The other authors describedsupramontane and montane sycamore communities from the mountains of other European countries:Mikyška (1972) from Sudets; Ellenberg, Klötzli from the Alps (1972); Wallnöfer et al. (1993) from the Alpsas a relict communities of previous periods, when the climate was more humid; Vacek et al. (2003) fromthe Czech Republic (Orlické hory); and Vukelič et al. (2008) from Croatia.

55

Acta Botanica Universitatis Comenianae Vol. 46, 2011

• Corresponding Author: Danica Černušáková; dcernusakova@fns.uniba.sk

Material and Methods

The phytocoenological relevés were performed according to the Zürich-Montpellier school (Braun-Blanquet 1951). The age of the trees in relevés number 1–5 were estimated, while the age of the treesin relevés 6-11 were determined according to the forestry maps from Oravice forestry district.Nomenclature of vascular plants follows Marhold et al. (1998); the nomenclature of syntaxa is takenfrom Wallnöfer et al. (1993). Diagnostic species of suballiance Aceri-Fagenion are assigned accordingto Husová (2000).

Eco-analysis follows the indicative method of Ellenberg (1978, 1991) with the update of eco-indicators (light, temperature, continentality, soil humidity, pH reaction, nitrogen content) published byKarrer, Killian (1990). For each eco-indicator the spectrum of weighted averages was calculated basedon the scoring of each species present in the association (Jurko 1983). This scoring, also known as thesignificance, xi of the i-th species in relevé, was evaluated as the abundance (transformed to numericalscale) multiplied by the constancy from the phytocoenological tables.

The significance was also used in the calculation of the following indices related to speciesdiversity of the herb layer (E1): Hill’s diversity index N2 (Hill 1973), Shannon’s diversity index H(Shannon, Weaver 1949), Pielou’s equitability index E (Pielou 1966) and Simpson’s index of theconcentration of dominance c (Simpson 1949), using the modifications for the species α-diversity assuggested by Jurko (1983). The explicit formulas for these indices are as follows (M is the total numberof species in the relevé):

The method (look up tables, algorithm of the indicative method of Ellenberg and calculation ofdiversity indices) is implemented in a simple way in the collection of MS-Excel worksheets and isavailable from the author upon request.

Study area. – Osobitá massif (1637 m) is located in the northwestern part of the Západné TatryMts. that are part of the Tatra National Park; since 1974, it is the National Nature Reserve (NNR).Osobitá is closed to public since 1988, so human activities are reduced to minimum. Most part of theOsobitá massif is built of limestone and dolomite from middle trias, with karstic relief, but southernand southeastern part is built from quartzites. They are formed to saddles and depressions (the name ofthe ridge Zadná Kremená stems from the quartzite bedrock).

The soil types in the area are tangled and leach rendzinas, less frequent are cambi-soils which aredeeper (with an average depth 40–100 cm) and richer in nutrients. Higher moisture of soil is one ofcondition for the development of these communities since it affects the speed of the humification.

56

According to Konček (1980), the climate is cold with an average annual temperature 4–6 °C. Thewarmest month is July with average temperature 14–15 °C. One of conditions for rich growth of theherb-containing sycamore-beech forests is high humidity. Total annual precipitation ranges between1200–1400 mm.

Location of relevés

1. Under Zadná Kremená, 800 m, exposition SW, inclination 5°, cover: E3 85 %, E2 5 %, E1 60 %, E0 2 %, age of forest 80,height 15 m, thickness 20 cm, 18. 7. 2006.

2. Zadná Kremená, 1250 m, exposition SW, inclination 10°, cover: E3 60 %, E2 5 %, E1 95 %, E0 0 %, age of forest 80,height 15 m, thickness 20 cm, 18. 7. 2006.

3. Zadná Kremená, 1300 m, exposition SW, inclination 20°, cover: E3 75 %, E2 2 %, E1 95 %, E0 10 %, age of forest 80,height 25 cm, thickness 50 cm, 18. 7. 2006.

4. Zadná Kremenná, 1100 m, exposition SW, inclination 30°, cover: E3 65 %, E2 1 %, E1 55 %, E0 1 %, age of forest 120,height 35 m, thickness 65 cm, 18. 7. 2006.

5. Zadná Kremená 1000 m, exposition SW, inclination 5°, cover: E3 75 %, E2 4 %, E1 80 %, E0 0 %, age of forest 100,height 20 m, thickness 35 cm, 18. 7. 2006.

6. Raganov žľab, right hand side of the creek, 1085 m, exposition NW, inclination 40°, cover: E3 75 %, E2 0 %, E1 95 %,E0 5 %, age of forest 100, height 20 m, thickness 30 cm, 1980.

7. Raganov žľab, right hand side of the creek, 1200 m, exposition NW, inclination 45°, total, cover: E3 75 %, E2 5 %, E1 95 %, E0 5 %, age of forest 100, height 25 m, thickness 30 cm, 1980.

8. Raganov žľab, 1260 m, exposition NNW, inclination 45–50°, cover: E3 60 %, E2 0 %, E1 95 %, E0 0 %, age of forest100, height 20 m, thickness 35 cm, 1980.

9. Široký žľab, 1200 m, exposition NW, inclination 35°, cover: E3 70 %, E2 0 %, E1 100 %, E0 0 %, age of forest 90, height20 m, thickness 30 cm, 1980.

10. Under Radové skaly (rocks), 1250 m, exposition N, inclination 45°, cover: E3 70 %, E2 0 %, E1 95 %, E0 15 %, age offorest 100, height 35 m, thickness 35 cm, 1980.

11. Under Radové skaly (rocks), 1280 m, exposition N, inclination 5°, cover: E3 65 %, E2 15 %, E1 60 %, E0 3 %, age offorest 80–100, height 15–20 cm, thickness 70–80 cm, 1980.

Results and Discussion

Phytocoenology

Phytosociological relevés are presented in Tab. 1. The montane sycamore-beech forests fromOsobitá belong to the community Cortuso-Fagetum (Klika 1927) Fajmonová 1982 (syn: Fagetumcarpaticum (calcicolum) Cortusae Klika 1927; Fagetum montanum carpaticum cortusae Klika 1936;Abieto-Fagetum carpaticum cortusae Klika 1946; Abieto-Fagetum adanostyletosum Fatrae Klika1949), suballiance Acerenion pseudoplatani Oberdorfer 1957, em. Husová in Moravec et al. (1982),alliance Fagion Luquet 1926.

Montane sycamore-beech forests grow on steep slopes with varying exposition, with the gradientsaround 40°. These communities prefer shady areas. This preference manifests itself also in the largernumber of species in shady stands with humid, nutritious, alkaline or weakly acid soil.

The crucial factor for the development of this plant community air- and soil humidity as indicatedin the floristic composition of the undergrowth (E1 level).

Fagus sylvatica and Acer pseudoplatanus are presented in the tree layer with larger coverage.Hygrophilous species, e.g., Adenostyles alliariae, Petasites albus, Stellaria nemorum andChrysosplenium alternifolium, which are diagnostic species of the suballiance Acerion, prevail in theherbs layer. They belong to species that are frequently presented in herb-rich spruce forest and grow inthe same altitude.

Association Cortuso-Fagetum is determinated by a few calcifilous species that are not presented atassociation Aceri-Fagetum: Polystichum lonchitis, Cortusa matthiolii, Cirsium erisithales andAsplenium viride.

57

The cover of the tree layer (E3) is lower (60–75 %, seldom 85 %) in comparison with theneighboring spruce or beech stands. In the tree layer, there are Fagus sylvatica, Picea abies, Acerpseudoplatanus, less frequently Abies alba, Sorbus aucuparia.

The shrub layer (E2) is weakly developed (2–4 %) and built mostly by juvenile individuals of thespecies from the layer and Ribes alpinum and Lonicera nigra.

The herb layer (E1) is made up of hygrophilous species and also nitrophilous ones: Geraniumrobertianum, Mercurialis perennis, Urtica dioica and Impatiens noli-tangere. Several other species arepresent – various beech species: Pulmonaria obscura, Paris quadrifolia, Actea spicata and also someferns as Dryopteris filix-mas, D. carthusiana, Polystichum lonchitis, Athyrium filix-femina. The herbfloor is distinctively multi-layered, especially remarkable are Adenostyles alliariae, Lunaria rediviva,Cicerbita alpina and Senecio ovatus.

Old dead trees remain in place and are an important source of nutrients for other species. It shouldbe noted that the positions of these stands are rather extreme; this is manifested by the reduced heightof the trees (around 20 m) and also by their thickness (often around 30 cm). According to the forestrymaps from Oravice forestry district the average age of these trees is 100 years.

Table 1 contains relevés from two different parts of Osobitá. Relevés numbered 1–5 (recorded in2006) in Zadná Kremená are more species-rich in the herb layer because the soil is deeper andnutritious. The relevés with numbers 6–11 recorded in 1980/1981 originate from Radové skaly with theshallow soils – rendzinas; here the bedrock often emerges to the surface.

Beech-sycamore forests of Osobitá are well preserved and, according to the age, shape and richundergrowth, they look healthy. Interestingly, in other regions of central European Carpathians theseforests are on decline. There are several factors contributing to this apparent discrepancy. 1. Osobitá massif is rather inaccessible and not suitable for commercial forestry (no regular work-outand planting). Limited management and forestry activity, for instance, no preference of spruce or fir,enables the slowly growing species to reproduce and sustain the competition of other species. For beechand sycamore this is an important factor, because their seeds are heavier then seeds of spruce or fir andthey spread slowly. Although juvenile sycamore grows faster than spruce, this is reversed for oldertrees. According to Heike (1978) 10 year old spruce grows already faster than beech. 2. The favorable climatic conditions, especially wind’s direction and relative humidity, support thedevelopment and prosperity of this community. Osobitá massif is not screened by higher mountainsfrom the north, therefore, the oceanic influence from the north and northwest is rather large with richand frequent precipitation (rain, snow) as well as fogs. 3. The declaration of the highest level of protection in 1988 was important factor for the build-up andrestoration of the forest and subalpine vegetation and flora. Despite the strict official protection, theterritory was not kept from some forest-management activities that we could observe during our fieldwork in 1980 and 2006, namely cutting down the old trees (mainly spruce and fir) in some easilyaccessible localities.

Ecoanalysis

Ecoanalysis is presented in Fig. 1 as a compact spectrum of relative significances of species presentin the community with respect to the Ellenberg’s ecoindices. It confirms the conclusions arising fromthe phytocoenology analysis.

The continentality has the centre of gravity between 3 and 4, i.e. oceanic to suboceanic species. Soilmoisture factor exhibits a broad peak around 6, indicating the preference of mesophilous tohygrophilous species, while nitrogen content peaks at 7, indicating the prevalence of seminitratophilousspecies and the temperature peak is spread almost equally between cryophilous and mesothermophilousspecies. The remaining two ecofactors (light and soil pH) exhibit multimodal spectrum of weightedaverages of relative species significance.

Diversity indices matrix is presented in Tab. 2. The main purpose of this part of eco-analysis is tomonitor the integrity of the relevés, i.e. the consistency of our results. It is interesting that the diversity

58

matrix splits into two blocks related to two groups of geographical locations: Zadná Kremenná valley(relevés 1–5) and glens in Raganov žľab/Radové skaly (relevés 6–11). The relevés from ZadnáKremenná are floristically slightly richer compared to the second group, as suggested by the Hill’s andalso Shannon’s indices. Otherwise the data in Table 2 indicate that the diversity is similar within thetwo mentioned groups. There are two exceptions – relevés no. 2 and 7, both seem to be taken frompartly disturbed stands, because both the diversity (either N2 or H) and the equitability indices are lowerthan in the remaining relevés. The values of Simpson’s index document that there are no extraordinarydominant individuals among the species recorded in Tab 1.

Summing up, the phytosociological data from Osobitá massif can contribute to the optimal strategyin the forestry management. Results from Osobitá can be utilized also in the studies of other protectedareas or for the restoration of disturbed/damaged forests in northern Slovakia. For instance, theevaluation of our results allows one to arrive to few useful hints for strategic thinking and landscapeplanning.

Although beech is slowly growing tree, it is more resistant against wind uprooting. In addition, it is more immune against air pollution and acid rains. It is necessary to keep and support the beech-sycamore communities in the harsh localities or in the localities, where the mixed forests representmore durable stands than pure coniferous ones. In the long run, the slower growing mixed forests arealso economically more profitable than fast growing conifers. One of the important roles of montanecommunities – protection of the soil against ablation and regulation of the water cycle – can besupported only by healthy, well-balanced forests.

Tab. 1. Phytocoenological relevés from Osobitá massifCortuso-Fagetum Klika (1927) Fajmonová 1982

59

Relevé numberNumber of species in relevé

E3

Picea abiesFagus sylvaticaAcer pseudoplatanusAbies albaSorbus aucupariaE2

Fagus sylvaticaLonicera nigraPicea abiesAcer pseudoplatanusSorbus aucupariaRibes alpinumE3

Diagnostic species of suballiance AcerenionAdenostyles alliariaeCicerbita alpinaPetasites albusThalictrum aquilegiifoliumRanunculus platanifoliusAconitum firmumChaerophyllum hirsutumAnother speciesOxalis acetosellaDryopteris filix-masMercurialis perenniGeranium robertianumPolystichum lonchitis

147

33.1.

1..1..

2+.1...

212+.

234

32.1.

1..1..

.

.

.1...

.

.2..

357

22..2

.++..+

31.11..

3231.

430

312..

+...+.

21..1..

22212

529

122..

++....

31..1..

333..

634

112+.

.+....

+.2+.+.

+1.2+

731

2r2.+

.

.r...

+.1....

+1.1+

826

2.2.1

.

.

.

.

.

.

1r2....

.1..+

926

1+2..

.

.

.

.

.

.

1.1....

11111

1027

+32+r

.

.

.

.

.

.

+....++

1.321

1136

13+1.

.

.+...

+...+..

211.1

C

VVIVIIIIII

IIIIIIIIIIIII

VIIIIIIIIIIIIIII

VVIVIVIV

Tab. 1. Continuation

60

Relevé numberNumber of species in relevé

Mycelis muralisStellaria nemorumChrysosplenium alternifoliumMyosotis sylvaticaUrtica dioicaMilium effusumCalamagrostis villosaImpatiens noli-tangereDryopteris carthusianaSenecio ovatusAthyrium filix-feminaGentiana asclepiadeaPulmonaria obscuraAcer pseudoplatanus juv.Luzula luzuloidesParis quadrifoliaEpilobium montanumActaea spicataLunaria redivivaMelampyrum sylvaticumHomogyne alpinaPrenathes purpureaSoldanella carpaticaHieracium murorumAbies alba juv.Luzula sylvaticaVaccinium myrtillusCardamine enneaphyllosRanunculus lanuginosusCardamine flexuosaGeranium sylvaticumAsarum europaeumPolygonatum vericillatumPhyteuma spicatumSorbus aucuparia juv.Veratrum albumCardaminopsis halleri subsp. tatricaRubus idaeusMaianthemum bifoliumPicea abiesHordelymus europaeusLilium martagonPrimula elatiorBellidiastrum micheliiCortusa matthioliCirsium erisithalesAsplenium virideGaleobdolon montanumE0

Brachythecium rivulareFissidens taxifolius .Conocephalum conicum

147

+....1.211112.++2..22.22221....2.1.++1.+......

.

.

.

234

.

.

.

.

.

.4....1.......3...1+.1......+..1.+..+.1.2..

.2.

357

1..+.13.2212.+21.1.22122.211....22.11+1..+322+..

2..

430

+....1..2.1...1...2.222...12....211.........1.1.

.

.

.

529

+.....2.2121.111.1..221.2.........1..1..1.......

.1.

634

.3+1++.2....r.r.+.+........+++r+...............+

.

.+

731

.411++.2+.......+........r..1+++...1....r.......

+..

826

.4121+.1....++.r+3.......+..1++....1....+.......

.

.

.

926

+2212+.3..+.1..r............1++r................

.

.

.

1027

1++11..1.1..1+....3+...................r........

2.+

1136

++1.+.+..1.+..r.r.1+..1r+......+....+..r..r...++

.I.

C

IVIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIII

IIII

Species in one relevéAnthoxanthum odoratum 1 (2), Anthyllis vulneraria + (2), Aquilegia vulgaris + (2), Aruncus vulgaris 1 (1), Astrantia major2 (2), Cardamine trifolia 1 (5), Carduus glaucinus 1 (2), Carex alba 1 (2), Carex sylvatica 1 (1), Crepis paludosa r (9),Dactylis glomerata r (8), Digitalis grandiflora 1 (3), Doronicum austriacum + (7), Epipactis atrorubens 1 (2), Eupatoriumcannabinum 1 (3), Fagus sylvatica juv.1 (1), Fragaria vesca + (11), Galium odoratum + (1), Galium schultesii 1 (2),Gymnocarpium dryopteris 1 (3), Heracleum sphondylium + (10), Hieracium sabaudum + (3), Hylotelephium maximum + (7),Laserpitium latifolium 2 (2), Linum catharticum 1 (2), Listera cordata 1 (3), Lonicera nigra 1 (1) juv., Luzula pilosa 2 (11),Lysimachia nemorum 1 (1), Melandrium rubrum + (3), Moneses uniflora + (2), Phegopteris connectilis 1 (1), Phyteumaorbiculare 1 (2), Pimpinella saxifraga 2 (2), Sanicula europaea 3 (1), Scrophularia scopolii + (7), Stachys sylvatica + (6),Thymus pulcherrimus 1 (2), Vaccinium vitis-idaea r (1), Valeriana simplicifolia 1 (2), Valeriana tripteris 2 (2), Veronicamontana 1 (7), Viola reichenbachiana + (1).

EoCtenidium molluscum + (11), Dicranum scoparium 1 (1), Metzgeria conjugata + (11), Plagiomnium cuspidatum 1 (6),Plagiothecium denticulatum 2 (6), Plagiomnium medium + (9), Polytrichum sp. 2 (3), Rhizomnium punctatum + (11).

Tab. 2. Diversity indices for the community Cortuso-Fagetum

Fig. 1. Ecoanalysis of the community Cortuso-Fagetum displaying the spectrum of weighted averages of six ecofactors

Acknowledgements

The help and numerous advices from Ján Bistar from TANAP during our field work in 2006 areacknowledged.

61

Index/Relevé 1 2 3 4 5 6 7 8 9 10

N2 18.7 5.0 13.7 15.2 8.0 5.1 3.5 5.0 9.4 5.2

H 3.0 1.7 3.3 3.4 3.2 2.8 2.3 2.7 3.4 3.0

E 0.6 0.3 0.6 0.7 0.7 0.6 0.5 0.6 0.8 0.7

c 0.1 0.2 0.1 0.1 0.1 0.2 0.3 0.2 0.1 0.2

40

35

30

25

20

15

10

5

0

Rela

tive s

pecie

s s

ignific

ance (

%)

Ellenberg Ecoindex

Spectrum of weighte averages

0 1 2 3 4 5 6 7 8 9

Light

Temp.

Contin.

Soil moist.

Soil pH

Nitrogen

References

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Wallnöfer, S., Mucina, L., Grass V., 1993: Querco-Fagetea, Űbersicht der Vegetationseinheiten. In: Mucina, L., Grabherr,G., Wallnöfer, S. (eds.), Die Pflanzengesellschaften Österreichs. Teil III, Wälder und Gebüsche, p. 85-212, Gustav FischerVerlag Jena, Stuttgart, New York.

Abstrakt

Príspevok je zameraný na spoločenstvá horských javorovo bukových lesov z masívu Osobitej v Západných Tatrách. Lesné spoločenstvá patria do asociácie Cortusao-Fagetum (Klika 1927)Fajmonová 1982, na študovanom území sa vyskytujú mozaikovito a zasahujú až po hornú hranicu lesa.Pre svoj vývoj potrebujú vyššiu vzdušnú aj pôdnu vlhkosť a na humus bohatšiu pôdu. Spoločenstvájavorovo bukových lesov tu netvoria súvislé porasty. Do ich vývoja zasiahol človek len minimálne,lebo sa nachádzajú na menej prístupných miestach. Tiež neboli narušené prírodnými katastrofami, a preto je potrebné aj naďalej sledovať vývoj týchto spoločenstiev, aby sa získané poznatky dali využiťpre zachovanie a ochranu podobných ekosystémov.

Danica Černušáková: Horské javorovo-bukové lesy masívu Osobitej (Západné Tatry) – fyto -cenologická a ekologická analýza.

62

USE OF DESCRIPTIVE STATISTICS IN PHYTOSOCIOLOGY

Peter Kučera•

Comenius University in Bratislava, Botanical Garden, Detached unit in Blatnica 315, 038 15 Blatnica, Slovakia

Received 13 December 2010; Received in revised form 6 October; Accepted 9 October 2011

Abstract

In terms of statistical methodics, use of the arithmetic mean is limited to ratio and interval scale data. Though, it isused within phytosociology most frequently for evaluation of the “average species cover” computed from theBraun-Blanquet cover-abundance scale, which is of ordinal character. Instead, application of more appropriatedescriptive statistics is proposed here. The paper summarizes basic facts on statistical properties of the Braun--Blanquet cover-abundance scale and relevant descriptive statistics. The use of descriptive statistics isdemonstrated on example of four communities of the order Athyrio filicis-feminae-Piceetalia Hadač ex Hadač etal. 1969.

Key words: arithmetic mean, biostatistics, measures of central tendency, measures of dispersion, ordinal scale

Introduction

Statistics, as the scientific study of data describing natural variation (Sokal, Rohlf 1997), is used inphytosociology in various statistical techniques spanning from determination of species constancy andaverage species cover to advanced clustering or ordination. However, theoretical background ofstatistics is repeatedly neglected for a long time. This is clearly visible for example in processing ofBraun-Blanquet cover-abundance data with use of the arithmetic mean (Fajmonová 1983, 1986;Jarolímek et al. 1997; Kliment 1997, 2005, 2010, Valachovič et al. 2001; Bernátová et al. 2003;Krajčiová-Šibíková et al. 2005; Šibík et al. 2005; Dúbravcová, Šibík 2006; Kliment et al. 2007;Jarolímek, Kliment 2008; Kochjarová et al. 2010). Topercer (1999) has already commented using ofmeasures of central tendency in phytosociology and using of ratio vs. ordinal scale as well, however,his paper was hardly noticed. Podani (2006) has also discussed the same topic, but his contribution ismore concentrated towards advanced, clustering and ordination statistical methods. The presentedpaper is an attempt to give an abbreviated view “inside” the very basic statistics with the aim tounderline important facts on use of descriptive statistics and relevant measurement scales as well.

Material and methods

Use of special biostatistical (biometrical) terms follows mostly Zar (1996), partly Sokal, Rohlf(1997). The terms “cover-abundance” and “cover-abundance scale” are taken from handbook ofWesthoff, van den Maarel (1973).

Phytosociological relevés were obtained from the Central database (2006) or inserted into databasevia the program Turboveg (Hennekens 2005). Then, they were processed by programs JUICE (Tichý2006) and SYN-TAX 2000 (Podani 2001a) to perform an ordinal cluster analysis of relevés: the

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Acta Botanica Universitatis Comenianae Vol. 46, 2011

• Corresponding Author: Peter Kučera; peter.kucera@rec.uniba.sk

coefficient Podani’s discordance was used (see Podani 2001b). Selection of relevés is commented byKučera (2007); syntaxonomical classification corresponds to that study, too. Source relevé groups(Tab. 2: community Sesleria-Pinus cembra and associations Seslerio variae-Piceetum, Cortuso-Piceetum, Polysticho lonchidis-Piceetum) included respectively 5, 13, 31 and 11 phytosociologicalrelevés. After a file export for computing of statistics of respective syntaxon, values for median, modeetc. were re-coded from original Braun-Blanquet cover-abundance values: r = 1, + = 2, 1 = 3, 2 = 4, 3 = 5, 4 = 6, 5 = 7, value “0” signifies absence of species in particular relevé. Statistics were computedby Microsoft® Excell 2000.

Species in Tab. 2 are ordered simply by their constancy within all syntaxa (details in Kučera 2007).Species of constancy less than 30 % are omitted. Constancy values are rounded to one decimal place.

Notes on use of statistics in phytosociology

Measurement scales

In general, statistical data can be recorded in four scales: ratio, interval, ordinal and nominal (Zar1996). The cardinal requirement for working with statistical data is to know the differences betweenthe scales. It is advisable to consider description and characteristics of the scales and the descriptivestatistics (see below) as well, as given in handbooks concerned on biostatistics or biometry, which dealalso with some basic theory of the science, prevailing with user-friendly language for non-mathematicians, e.g. Bablo (1966), Bauer et al. (1967), Bakytová et al. (1975), Komenda, Klementa(1981), Jedlička (1984), Lukasová, Šarmancová (1985), Hebák, Hustopecký (1987), Clauss, Ebner(1988), Bakytová et al. (1996), Lepš (1996), Zar (1996), Sokal, Rohlf (1997), Marhold, Suda (2002).

Within a chosen phytosociological relevé plot in the field, plant species as well as vegetation layersoccupy exact cover values expressed in square metres or centimetres. Such exact data could bestatistically treated as data of the ratio scale. Mentioned data type is in the Braun-Blanquet approach(cf. Westhoff, van den Maarel 1973; Moravec 1994) estimated primarily in per cents. [Percentages arethe type of a ratio, which expresses the relation between two variables as a single value. A ratio is oneof derived variables (Jedlička 1984; Sokal, Rohlf 1997).] Nevertheless, usual visual field estimation ofspecies cover is definitely not capable to express the cover in per cents as well. Concerning the usualpurpose of phytosociological relevés, namely presentation of composition and approximate structure ofvegetation, it would also be (in the most cases) useless vaste of time requiring expensive technicalequipment. Instead, cover and abundance of species are recorded in useful Braun-Blanquet cover--abundance scale (Braun-Blanquet 1951: 60–61), with certain limitations resulting from visualestimation. However, this scale is example of the ordinal scale (cf. Kent, Coker 1996; Podani 2006).Species cover-abundance can be recorded also in the nominal scale: as presence or absence data of a particular species.

Basic feature of the ordinal scale is that it allows only to arbitrate about the order between everytwo variates of dataset (X1 < X2), while the ratio scale allows to express the difference between twovariates, and also the ratio between them. “Ordinal scale data contain and convey less information thanratio or interval data, for only relative magnitudes are known. Consequently, quantitative comparisonsare impossible…” (Zar 1996). For example: Any species cover between 25 and 50 % of relevé area isrecorded in the Braun-Blanquet scale as a degree 3 (Braun-Blanquet 1951). As a rule, once the real fielddata (= exact values in square meters, expressed in ratio to size of relevé plot) are recorded in theordinal scale, they can not be converted back to the ratio scale. Degree of species cover-abundance 3could be expressed alternatively only in the nominal scale: as value 1 – the species is present in therelevé area. Table 1 brings a comparison of the three scales: ratio, ordinal and nominal. Converting ofdata accuracy is to be applied only from left side of the table to its right side, in no case in revertdirection.

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Tab. 1. Comparison of expression of species cover-abundancevalues within the ratio, ordinal and nominal scale.

Notes to Tab. 1:*1 Numbers 0–100 represent the total cover of an individual species expressed in per cents. The maximum of the Braun-

-Blanquet cover-abundance scale (recorded as degree 5, Braun-Blanquet 1951: 61) is total cover of 100 %, which is definitenumber. It is needful to have defined also another degrees in this manner as, for example, species cover of 75 % can not beassigned to more than one degree (cf. Tüxen, Elleberg 1937; Braun-Blanquet 1951, 1964; Westhoff, van den Maarel 1973;Moravec 1994 vs. Kent, Coker 1996). Consequently, cover-abundance degree 5 should be defined as any value of cover frommore than 75 % up to 100 % accurately: mathematically expressed with a half-open interval “(75 %; 100 %>”, adequatelyother major degrees: 4 – (50 %; 75 %>; 3 – (25 %; 50 %>; 2 – (5 %; 25 %>. Accordingly, the upper limit of the cover-abundance degree 1 is set here to maximum cover of 5 % precisely: in this manner, degrees 2 to 5 of the original Braun-Blanquet cover-abundance scale are defined analogously. Values 0.1 %, 5.1 %, 25.1 % etc. are selected only as an example(it could be also “0.01 %, 5.01 %…”).

*2 The ordinal scale is represented by the phytosociological Braun-Blanquet cover-abundance scale. It was refined byBarkman et al. (1964), however, only degrees 2m, 2a, 2b became into wider use (cf. Westhof, van den Maarel 1973).Numerals 0–9 are used “to code” the scale for computing purposes. Nevertheless, such a re-coding of degrees could not belabelled as “ordinal data transformation” (cf. Westhoff, van den Maarel 1973: 936, Tab. II; Krahulec 1994: 130), as theoriginal scale is already of ordinal character.

The Braun-Blanquet cover-abundance scale (Braun-Blanquet 1951) [and other derivated scales, e.g. Zlatník (1953); formore scales cf. van den Maarel (1979: Tab. 2)] is based on the cover scale (Braun-Blanquet 1951: 59). In this scale, the degree1 means given species cover up to 5 %. Transferred in the cover-abundance scale, this degree was divided into three degrees:r , + and 1 (compare the Braun-Blanquet abundance scale!: Braun-Blanquet 1951). Originally, latter three degrees weredefined only verbally, not quantitatively. For this reason, no quantitative values can be assigned to any of them (except ofpotential brand new definition of the Braun-Blanquet cover-abundance scale; however, such proposal should be consideredcarefully). According to their definitions (see Braun-Blanquet 1951: 60, Barkman et al. 1964), these three degrees could onlybe ordered in array “r, +, 1”.

*3 The nominal scale is represented here by its special case: the binary scale, as only values 0 (absence of a species) and1 (presence of a species) are relevant.

Descriptive statistics

Quantities concentrating the information about a sample and stating on sample characteristics arecalled statistics, e.g. arithmetic mean (Jedlička 1984). Statistics (Zar 1996) or descriptive statistics(Sokal, Rohlf 1997) are divided into two main groups: (1) measures of central tendency [statistics oflocation: Sokal, Rohlf (1997)] – arithmetic mean (usually: mean, average), geometric mean, harmonicmean, median, mode, quartiles and other quantiles and (2) measures of dispersion [statistics ofdispersion: Sokal, Rohlf (1997)] – range, interquartile range, semi-interquartile range, variance,standard deviation, coefficient of variation. In addition, third group of measures – symmetry(skewness) and kurtosis – is distinguished (cf. Jedlička 1984: 28). Basic information on range ofvariates is provided by the minimum (Xmin) – the lowest value of dataset, and, the maximum (Xmax) –the highest value of dataset.

The arithmetic mean is a descriptive statistics familiar from common use. It equals to sum of allthe individual observations or items of a sample divided by the number of items in the sample (Sokal,Rohlf 1997). The arithmetic mean is the most commonly used statistics in phytosociology, too. It isused for presentation of “average cover values” of particular species in synoptic tables, e.g. Jarolímeket al. (1997) and later published parts of the series Plant communities of Slovakia; cf. Introduction).

65

ratio scale*1

0

0.1–5.0

05.1–12.5

12.6–25.0

25.1–50.0

50.1–75.0

75.1–100

ordinal scale*2

0

1 (r), 2 (+), 3 (1), 4 (2m)

5 (2a)

6 (2b)

7 (3)

8 (4)

9 (5)

nominal scale*3

0

1

By contrast, Zar (1996) specifies that the arithmetic mean is applicable only to interval or ratio scaledata. Only in these two scales is allowed addition and subtraction of values, in the latter one alsomultiplication and division. That is why the arithmetic mean can not be used, when phytosociologicaldata on ordinal scale are considered: nor as simple value, neither within various methods using it andhandling the data as they were real values.

Under the rules, given species cover of the cover-abundance degree 3 (from the example above)can not be substituted for computing purposes subsequently by value 38 [%, resp. 37.5 %], which is meant as the arithmetic mean of only two marginal values of cover range 25–50 %, as for exampleprograms JUICE (Tichý 2006) or Turboveg (Hennekens 2005) do in specific cases. One should realizethe difference between computerised coding of field data, however required and necessary and actualcomputations for data analysis. Subsequent substitution of original ordinal data by percentage values isby no means related to real cover values in the sample. From the other point of view, this would implythat all species with original cover-abundance degree 3 in the given sample of relevés have had in thefield real cover value 38 % [or the arithmetic mean of real cover values within processed relevés isalways 38 %]. Also the Braun-Blanquet cover-abundance values r, +, 1 can not be converted torespective values 1 %, 2 %, 3 %, [2m (cf. Barkman et al. 1964) to 4 %] (cf. JUICE) or + to 0.1 %, 1 to2.5 % etc. [cf. Braun-Blanquet 1948: 268], for they are ordinal values. Using the proper, ordinalstatistical methods, e.g. Podani (2006), there is actually no need of such conversion at all.

Tradition of subsequent “reverse conversions” of cover-abundance values to percentages isprobably based on early work of Tüxen, Ellenberg (1937). Their paper included several proposals ofusing mathematics for phytosociological purposes. Although the authors did not consider the statisticalcorrectness of their method of expressing the “mittlere Gruppenmenge” (Tüxen, Ellenberg 1937: 173;see Appendix 1), recent phytosociologists are not apologized with mistake of that authors and shouldhave to consider basic principles of using/processing the statistical data. Braun-Blanquet (1951: 109;1964: 52) followed Tüxen, Ellenberg (1937), but with some changes: to degree 2 was assigned therange of cover values 10–25 %, to degree 1 range 1–10 %. The corresponding “MittleresDeckungsprozent”-values were set to 17.5 % and 5.0 % respectively (cf. Braun-Blanquet et al. 1946sec. Braun-Blanquet 1964: 39).

The arithmetic mean is also generally used when processing of vascular plants species indicatorvalues of Ellenberg (1974) is needed (Majzlanová 1983, 1993; Šomšák et al. 1984; Majzlanová,Matejovič 1986; Černušáková 1994; Chomová 2002; Kanka 2001). Nevertheless, these are, just asBraun-Blanquet cover-abundance values, only codes of various degrees of some plant characteristics andnot real measurable values such as height of plant species or length of its leaves in centimetres, weightof an individual seed and so on. The same is to be said on some ecological and socioeconomic values ofplants of Jurko (1990): soil humidity, soil pH, nutrient value of soil etc. (remark that some Jurko’s valuesare data processed only with nominal scale: life forms, consistency of leaves, duration of leaves and soon). The Ellenberg’s ecological species values and some Jurko’s values are ordinal scale data.

Similarly as the arithmetic mean, some other measures (harmonic mean, variance, standarddeviation etc.) can not be used with Braun-Blanquet cover-abundance scale as they require performingof mathematical operations not compatible with ordinal scale, i.e. multiplication, division as well asaddition and subtraction.

The median is a value dividing the non-decreasing order of values within the sample into two equalhalves: the median of (ordered) dataset “1, 1, 1, 2, 3, 4, 4, 4, 6” is value 3. When a sample has evennumber of values (e.g. “2, 3, 3, 4, 4, 5”, there are two middle values – 3 and 4: then the median isconventionally defined as the midpoint between them (Kent, Coker 1997; Zar 1996; Sokal, Rohlf1997). In this case, it is the value 3.5. Unlike the arithmetic mean, the median can be used also withordinal scale data (Zar 1996). If the frequency distribution is symmetrical and unimodal, the value ofthe median belongs to the most frequent value in the sample (cf. Zar 1996). In such case, the medianis the suitable alternative for the arithmetic mean. Non-decreasing order of dataset values could bedivided also into fourths, so as one fourth of values is smaller than the first quartile (Q1), and one fourthlarger than the third quartile (Q3). The median is identical to the second quartile. A set of data could be

66

divided into more equal parts – values, which devide them are called generally quantiles. For moreadvanced comments to median and following measures, to their use and limitations, it is advisable toconsider specialized literature (e.g. Zar 1996; Sokal, Rohlf 1997).

The mode is the most frequently occurring value in a dataset: from the former example above it isthe value 4. A sample could have also two modes, if the frequency distribution is bimodal (Zar 1996).The mode can be used with ratio, ordinal and also nominal scale data.

The range (R) is the difference between the minimum and the maximum value of a sample. It givesthe range of margin values of a dataset. The range is also applicable to ordinal scale data, similarly asthe interquartile range (q) (Zar 1996). The latter measure is the difference between the first quartileand the third quartile. It express the range of 50 % of values (concentrated round the median), withoutvalues lower than first quartile and larger than third quartile. (Also quartile deviation could be used,which is a q-value divided by 2). Therefore the interquartile range is less sensitive to extremely largeand small values in the sample (Zar 1996).

Cover-abundance values of particular species are ordered in commonly publishedphytosociological tables according to the chosen order of phytosociological relevés, e.g. “2, 4, 4, 5, 1,2”. For determination of descriptive statistics, cover-abundance values of an individual species withinset of relevés have to be ordered to a non-decreasing order of values, e.g. “1, 2, 2, 4, 4, 5”: for exampleby the JUICE (Tichý 2006). This step could be skipped when statistics are computed automatically byMicrosoft® Excell 2000 from table files exported from the JUICE.

Characteristics of phytosociological relevés with descriptive statistics

Table 2 brings shortened statistical species evaluation of group of four plant communities (orderAthyrio filicis-feminae-Piceetalia Hadač ex Hadač et al. 1969), classified in the terms of Kučera (2007)as: (a) community Sesleria tatrae-Pinus cembra, (b) association Seslerio variae-Piceetum Fajmonová1978, (c) association Cortuso-Piceetum Fajmonová 1986 and (d) relevés preliminary grouped aroundthe association Polysticho lonchitidis-Piceetum W. Matuszkiewicz ex J. Matuszkiewicz 1977.Examples of statistical evaluation are given below in the form “species/column”, i.e. species namefollowed by the respective column label a, b, c, d.

Use of the measure minimum (Xmin) is self-explaining. Value “0” says that particular species areabsent in one sample relevé at least: Lonicera nigra (E1)/a–d, Oxalis acetosella/a–d. The larger valueof the minimum, the higher cover-abundance degrees has the particular species in the sample (plantcommunity, syntaxon): e.g. Picea abies (E3)/b, c, Pinus cembra (E3)/a, Vaccinium myrtillus/d,Calamagrostis varia/a. The minimum equivalent to Braun-Blanquet cover-abundance degrees (2), 3, 4,5 stand for a constantly dominant species with special role in vegetation structure and physiognomycorresponding to ecological factors.

The opposite measure, maximum (Xmax), is simply the largest value of species cover-abundance inthe dataset. The closer values of measures of central tendency and the minimum to the maximum value,the higher stability of species cover-abundance in the sample: Daphne mezereum (E1)/a, Salix silesiaca(E2, E1)/a, Homogyne alpina/b, Calamagrostis varia/a, Dicranum scoparium/a.

In given cases, the median is a suitable alternative for the arithmetic mean (see above). Medianvalue 0 implies that species has lower constancy than 50 %: Adenostyles alliariae/b, Cortusamatthioli/d, Dryopteris dilatata/b, c, d.

Within relevés of a sample (syntaxon), the mode estimates most frequently occurring cover--abundance value of a particular species (cf. Valeriana tripteris, Luzula sylvatica, Vaccinium myrtilus,Homogyne alpina). Consequently mode value 0 means that the most frequent group of cover--abundance values of particular species stand for absence of the species in the sample, although themedian could be greater than 0: Calamagrostis varia/c, Adenostyles alliariae/c, Gentianaasclepiadea/d, Primula elatior/b. Greater mode values indicate usually dominant species: Picea abies(E3)/b, c, Pinus cembra (E3)/a, Vaccinium myrtillus/d.

67

Value of the first quartile (Q1) greater than 0 indicates that species is present at least in 3/4 ofrelevés in given sample (syntaxon) and reversely. In the former case, the species belongs to the moststable components of specified plant community: Polygonatum verticillatum/a, b, Phyteumaspicatum/c, Clematis alpina/a, b, Calamagrostis arundinacea/b. [Similarly, non-zero value of thequantile Q20 would indicate species constancy class “V”, i.e. 81–100 %.] The greater first quartilevalue, the higher overall species cover in sample relevés.

If the value of the third quartile (Q3) is equal to 0, then a particular species is absent in as much as3/4 of sample relevés at least: Sorbus aucuparia (E3)/b, Picea abies (E2)/b, c, Cortusa matthioli/d,Cirsium erisithales/d, Sesleria tatrae/c, d [cf. the quantile Q80 for constancy class “I” (1–20 %)].Consequently, median and mode values are 0, too. This is the example of species with low constancywithin the dataset, only in special cases (as differential species) relevant in species composition of asyntaxon.

The greater the value of the range, the more variable (i.e. less stabilised) is cover-abundance of a particular species within a dataset (syntaxon): Picea abies (E3)/a, d, Pinus mugo (E2)/a,Calamagrostis varia/b, c, C. arundinacea/c, Mercurialis perennis/b. Value 0 signifies invariable degreeof species cover-abundance: Daphne mezereum (E1)/a, Salix silesiaca (E1)/a, Dicranum scoparium/a.

Among the statistics of dispersion, the interquartile range is less sensitive as the range to so-calledoutliers from both ends of cover-abundance variation range: Picea abies (E3)/a, d, Pinus mugo (E2)/b,Luzula sylvatica/b, Hieracium murorum/b, Vaccinium vitis-idaea/d, Leucanthemum rotundifolium/c,Hylocomium splendens/c, Mnium spinosum/b–d. In this manner, it gives more specific information onpattern of species cover-abundance.

Tab. 2. Statistics computed from the relevé set and constancy of species

68

E3

Picea abiesSorbus aucupariaPinus cembraBetula carpaticaSalix caprea

E2

Sorbus aucupariaPicea abiesPinus mugoSalix silesiacaPinus cembraBetula carpaticaLonicera nigra

E1

Daphne mezereumSorbus aucupariaLonicera nigraPicea abiesSalix silesiacaRibes petraeumPinus cembraAcer pseudoplatanus

Valeriana tripterisLuzula sylvaticaVaccinium myrtillusHomogyne alpinaCalamagrostis variaOxalis acetosellaPolygonatum verticillatumHieracium murorumVaccinium vitis-idaeaPhyteuma spicatumAdenostyles alliariaeCortusa matthioliClematis alpinaSoldanella hungarica

Xmin

a b c d

0 5 5 32 0 0 05 × × 00 × × 00 × × ×

2 × 0 02 0 0 02 0 0 01 × 0 00 × × 00 × × 00 × 0 0

1 0 0 02 2 0 00 0 0 00 × 0 02 × 0 01 × 0 ×0 × 0 0× 0 0 0

2 2 2 03 0 2 00 0 0 40 2 0 04 0 0 00 0 0 00 0 0 00 0 0 02 0 0 00 0 0 00 0 0 00 0 0 01 0 0 00 0 0 0

Xmax

a b c d

4 7 7 75 3 3 36 × × 34 × × 42 × × ×

4 × 3 34 2 3 36 1 3 22 × 2 33 × × 23 × × 33 × 3 2

1 3 3 23 3 3 32 2 2 33 × 2 32 × 3 22 × 4 ×2 × 1 2× 2 2 2

4 5 4 34 4 5 45 3 3 73 3 3 45 5 6 44 3 5 42 3 3 24 4 4 34 2 3 42 3 3 32 2 5 32 4 5 22 3 3 23 3 4 3

median

a b c d

3 6 6 63 0 2 06 × × 03 × × 00 × × ×

2 × 0 04 0 0 04 0 0 02 × 0 03 × × 01 × × 01 × 0 0

1 2 2 02 3 0 20 2 0 00 × 0 22 × 0 02 × 0 ×2 × 0 0× 0 0 0

3 3 3 23 3 4 33 3 2 51 3 2 35 3 2 22 3 3 32 2 2 23 3 2 22 2 0 32 2 2 01 0 2 22 2 4 01 2 0 02 3 0 0

mode

a b c d

4 6 6 63 0 0 06 × × 00 × × 00 × × ×

2 × 0 04 0 0 04 0 0 02 × 0 03 × × 01 × × 00 × 0 0

1 2 2 02 3 0 20 2 0 00 × 0 22 × 0 02 × 0 ×2 × 0 0× 0 0 0

4 3 3 23 3 4 30 3 2 50 3 3 25 3 0 20 3 3 32 3 2 24 3 3 02 2 0 32 3 2 02 0 0 22 0 4 01 2 0 02 3 0 0

Q1

a b c d

3 6 6 53 0 0 05 × × 00 × × 00 × × ×

2 × 0 03 0 0 04 0 0 02 × 0 02 × × 01 × × 00 × 0 0

1 2 0 02 3 0 20 0 0 00 × 0 0.52 × 0 01 × 0 ×0 × 0 0× 0 0 0

3 3 2 13 3 3 20 2 2 50 2 2 25 2 0 10 2 2 1.50 2 2 03 3 2 02 2 0 2.52 0 2 01 0 0 0.51 0 4 01 2 0 02 3 0 0

Q3

a b c d

4 6 6 64 0 2 26 × × 33 × × 32 × × ×

3 × 2 24 0 0 24 0 0 02 × 0 03 × × 02 × × 02 × 0 0

1 2 2 13 3 0 31 2 1 1.52 × 0 22 × 1 02 × 0 ×2 × 0 1× 0 0 1.5

4 3 3 23 3 4 3.54 3 3 62 3 3 3.55 4 2.5 33 3 3 32 3 3 24 3 3 23 2 0 42 3 3 22 2 3.5 22 3 5 02 2 2 22 3 3 2

R

a b c d

4 2 2 43 3 3 31 × × 34 × × 42 × × ×

2 × 3 32 2 3 34 1 3 21 × 2 33 × × 23 × × 33 × 3 2

0 3 3 21 1 3 32 2 2 33 × 2 30 × 3 21 × 4 ×2 × 1 2× 2 2 2

2 3 2 31 4 3 45 3 3 33 1 3 41 5 6 44 3 5 42 3 3 24 4 4 32 2 3 42 3 3 32 2 5 32 4 5 21 3 3 23 3 4 3

q

a b c d

1 0 0 11 0 2 21 × × 33 × × 32 × × ×

1 × 2 21 0 0 20 0 0 00 × 0 01 × × 01 × × 02 × 0 0

0 0 2 11 0 0 11 2 1 1.52 × 0 1.50 × 1 01 × 0 ×2 × 0 1× 0 0 1.5

1 0 1 10 0 1 1.54 1 1 12 1 1 1.50 2 2.5 23 1 1 1.52 1 1 21 0 1 21 0 0 1.50 3 1 21 2 3.5 1.51 3 1 01 0 2 20 0 3 2

constancy (%)

a b c d

80.0 100.0 100.0 100.0100.0 23.1 54.8 36.4100.0 – – 45.4

60.0 – – 36.440.0 – – –

100.0 – 32.3 45.4100.0 23.1 19.4 36.4100.0 23.1 16.1 9.1100.0 – 16.1 9.1

80.0 – – 9.180.0 – – 9.160.0 – 3.2 18.2

100.0 92.3 67.7 45.4100.0 100.0 12.9 81.8

40.0 69.2 25.8 45.440.0 – 22.6 72.7

100.0 – 25.8 9.1100.0 – 16.1 –

60.0 – 3.2 36.4– 15.4 12.9 45.4

100.0 100.0 100.0 72.7100.0 84.6 100.0 81.8

60.0 92.3 83.9 100.060.0 100.0 80.6 90.9

100.0 84.6 54.8 72.760.0 76.9 83.9 81.860.0 92.3 83.9 63.680.0 84.6 80.6 54.5

100.0 84.6 12.9 90.980.0 69.2 80.6 45.480.0 46.2 58.1 72.780.0 61.5 93.5 18.2

100.0 76.9 35.5 36.480.0 84.6 38.7 45.4

Tab. 2. Continuation

69

Asplenium virideCirsium erisithalesPrenanthes purpureaAster bellidiastrumGeranium sylvaticumGentiana asclepiadeaSenecio nemorensis agg.Dryopteris dilatataMelampyrum sylvaticumMaianthemum bifoliumAstrantia majorPrimula elatiorPolystichum lonchitisGymnocarpium dryopterisGalium schultesiiAvenella flexuosaHeracleum sphondyliumSesleria albicansViola bifloraCrepis paludosaCardaminopsis arenosa agg.Calamagrostis arundinaceaVeratrum * lobelianumFragaria vescaMoneses unifloraPhyteuma orbiculareSwertia perennisMycelis muralisSesleria tatraeCarex *tatrorum (Zapał.)Pawł.Rubus saxatilisCystopteris montanaCrepis jacquiniiMercurialis perennisLeucanthemumrotundifoliumCystopteris fragilisGaleobdolon luteum agg.Huperzia selagoPoa alpinaPimpinella majorCampanula tatraeSolidago virgaureaLuzula luzuloidesCampanula cochleariifoliaChaerophyllum hirsutumCarex digitataDryopteris filix-masSoldanella carpaticaTanacetum clusiiRanunculus breyninusEpilobium montanumCarduus glaucinusHedysarum hedysaroidesPyrola rotundifoliaBartsia alpinaRanunculus platanifoliusSenecio subalpinusDentaria enneaphyllosTofieldia calyculataRumex alpestrisMyosotis sylvaticaLilium martagonGeum rivaleCicerbita alpinaCampanula rotundifolia agg.Rubus idaeusCarex ornithopodaRhodiola roseaSalix retusaSalix reticulataFestuca versicolorAndrosace chamaejasmeMoehringia muscosaRanunculus alpestris

Xmin

a b c d

0 0 0 00 0 0 00 0 0 00 0 0 00 0 0 0× 0 0 00 0 0 00 0 0 00 0 0 0× 0 0 00 0 0 0× 0 2 00 0 0 00 0 0 00 0 0 00 0 × 0× 0 0 0× 3 0 ×0 0 0 ×× 0 0 0× 0 0 0× 0 0 00 0 0 0× 0 0 00 0 × 00 0 × 00 0 0 0× 0 0 02 × 0 0

0 0 × ×× 0 0 00 × 0 00 0 × ×× 0 0 ×

0 0 0 ×1 × × 0× 0 0 00 × × 00 0 × ×× 0 0 ×1 × × ×× 0 0 00 0 × 0× 0 × 0× 0 0 0× 0 0 ×× 0 0 0× 0 0 0× 0 0 ×× 0 × ×× 0 0 0× 0 0 ×0 × × ×0 × × ×0 × × ×0 0 0 0× 0 0 0× 0 0 ×0 0 × ×0 × 0 ×× 0 0 0× 0 0 ×0 0 0 ×0 0 0 0× 0 × 00 0 × 0× 0 × ×0 × × ×0 × × ×0 × × ×0 × × ×0 × × ×0 × × ×0 × × ×

Xmax

a b c d

3 3 3 21 3 3 12 3 3 34 3 4 33 3 3 3× 3 3 32 3 4 32 2 3 54 3 3 3× 4 4 41 3 3 3× 3 4 32 3 3 23 3 4 31 3 3 21 2 × 4× 2 4 1× 6 3 ×2 3 4 ×× 3 3 2× 3 3 2× 4 6 31 2 3 2× 3 3 31 2 × 33 3 × 24 3 2 2× 3 3 24 × 2 2

4 3 × ×× 4 3 33 × 2 22 2 × ×× 6 4 ×

2 2 5 ×1 × × 2× 2 3 32 × × 22 2 × ×× 2 3 ×2 × × ×× 3 3 33 3 × 3× 3 × 2× 2 4 2× 3 3 ×× 2 3 2× 3 4 2× 3 3 ×× 3 × ×× 2 3 2× 2 2 ×2 × × ×3 × × ×2 × × ×3 1 3 2× 2 3 3× 3 5 ×2 3 × ×2 × 2 ×× 2 5 1× 2 2 ×2 2 3 ×1 2 3 3× 3 × 21 2 × 3× 3 × ×2 × × ×4 × × ×3 × × ×2 × × ×2 × × ×2 × × ×2 × × ×

median

a b c d

3 2 2 01 3 2 00 2 2 23 3 0 02 0 2 0× 2 2 10 0 2 01 0 0 02 0 0 0× 2 0 00 2 2 0× 2 3 00 2 1 02 0 0 00 0 2 00 1 × 2× 2 2 0× 5 2 ×0 2 3 ×× 0 2 0× 2 2 0× 3 0 00 0 1 0× 2 0 01 0 × 00 2 × 01 0 0 0× 2 0 03 × 0 0

2 2 × ×× 0 2 02 × 0 00 1 × ×× 2 2 ×

1 0 0 ×1 × × 0× 0 2 02 × × 01 0 × ×× 2 0 ×2 × × ×× 0 0 00 0 × 0× 2 × 0× 0 2 0× 3 0 ×× 0 0 0× 0 0 0× 0 2 ×× 3 × ×× 0 0 0× 1 0 ×2 × × ×2 × × ×2 × × ×0 0 0 0× 0 0 0× 0 2 ×0 2 × ×1 × 0 ×× 0 0 0× 0 0 ×0 0 0 ×0 0 0 0× 0 × 00 0 × 0× 2 × ×2 × × ×3 × × ×2 × × ×2 × × ×2 × × ×2 × × ×1 × × ×

mode

a b c d

3 0 2 01 3 2 00 2 0 20 3 0 00 0 2 0× 2 2 00 0 2 02 0 0 00 0 0 0× 2 0 00 2 2 0× 0 3 00 3 0 03 0 0 00 0 2 00 0 × 2× 2 2 0× 5 2 ×0 3 3 ×× 0 3 0× 3 2 0× 3 0 00 0 0 0× 2 0 01 0 × 00 2 × 01 0 0 0× 3 0 02 × 0 0

0 0 × ×× 0 0 00 × 0 00 2 × ×× 0 0 ×

2 0 0 ×1 × × 0× 0 2 02 × × 00 0 × ×× 2 0 ×2 × × ×× 0 0 00 0 × 0× 3 × 0× 0 2 0× 3 0 ×× 0 0 0× 0 0 0× 0 2 ×× 3 × ×× 0 0 0× 2 0 ×2 × × ×2 × × ×2 × × ×0 0 0 0× 0 0 0× 0 0 ×0 0 × ×0 × 0 ×× 0 0 0× 0 0 ×0 0 0 ×0 0 0 0× 0 × 00 0 × 0× 0 × ×2 × × ×0 × × ×0 × × ×2 × × ×2 × × ×2 × × ×0 × × ×

Q1

a b c d

2 0 0 00 2 2 00 2 0 10 3 0 00 0 1 0× 0 1 00 0 1 01 0 0 00 0 0 0× 2 0 00 2 0 0× 0 3 00 0 0 02 0 0 00 0 0 00 0 × 2× 2 1 0× 4 0 ×0 0 0 ×× 0 1.5 0× 0 0 0× 3 0 00 0 0 0× 2 0 00 0 × 00 2 × 01 0 0 0× 0 0 02 × 0 0

0 0 × ×× 0 0 00 × 0 00 1 × ×× 0 0 ×

0 0 0 ×1 × × 0× 0 0 01 × × 00 0 × ×× 0 0 ×1 × × ×× 0 0 00 0 × 0× 1 × 0× 0 0 0× 2 0 ×× 0 0 0× 0 0 0× 0 0 ×× 2 × ×× 0 0 0× 0 0 ×1 × × ×2 × × ×1 × × ×0 0 0 0× 0 0 0× 0 0 ×0 0 × ×0 × 0 ×× 0 0 0× 0 0 ×0 0 0 ×0 0 0 0× 0 × 00 0 × 0× 0 × ×0 × × ×0 × × ×0 × × ×0 × × ×0 × × ×0 × × ×0 × × ×

Q3

a b c d

3 3 2 21 3 3 00 3 2.5 2.54 3 2 02 2 2.5 1× 2 2 21 2 3 1.52 0 2 22 3 2 2× 3 2 20 3 2 0× 3 3 01 3 3 03 0 0 20 2 2.5 20 2 × 3× 2 2 0× 6 2 ×0 3 3 ×× 2 3 1.5× 3 2 0× 3 2.5 21 2 2 0.5× 2 1.5 1.51 2 × 12 3 × 03 2 0 0× 3 2 04 × 0 0

2 2 × ×× 2 3 12 × 0 11 2 × ×× 3 3 ×

2 2 0 ×1 × × 0× 0 2 12 × × 0.52 2 × ×× 2 2 ×2 × × ×× 2 2 0.50 2 × 2× 3 × 0× 0 2 0× 3 0 ×× 0 2 1× 2 3 0× 2 2 ×× 3 × ×× 0 2 1× 2 0 ×2 × × ×2 × × ×2 × × ×0 0 2 0× 0 2 0× 0 3 ×0 3 × ×2 × 0 ×× 0 2 0× 2 1.5 ×0 0 2 ×0 0 2 0× 3 × 00 0 × 1.5× 3 × ×2 × × 3 × × ×2 × × ×2 × × ×2 × × ×2 × × ×2 × × ×

R

a b c d

3 3 3 21 3 3 12 3 3 34 3 4 33 3 3 3× 3 3 32 3 4 32 2 3 54 3 3 3× 4 4 41 3 3 3× 3 2 32 3 3 23 3 4 31 3 3 21 2 × 4× 2 4 1× 3 3 ×2 3 4 ×× 3 3 2× 3 3 2× 4 6 31 2 3 2× 3 3 31 2 × 33 3 × 24 3 2 2× 3 3 22 × 2 2

4 3 × ×× 4 3 33 × 2 22 2 × ×× 6 4 ×

2 2 5 ×0 × × 2× 2 3 32 × × 22 2 × ×× 2 3 ×1 × × ×× 3 3 33 3 × 3× 3 × 2× 2 4 2× 3 3 ×× 2 3 2× 3 4 2× 3 3 ×× 3 × ×× 2 3 2× 2 2 ×2 0 × ×× 3 × ××2 × × ×3 1 3 2× 2 3 3× 3 5 ×2 3 × ×2 × 2 ×× 2 5 1× 2 2 ×2 2 3 ×1 2 3 3× 3 × 21 2 × 3× 3 × ×2 × × ×4 × × ×3 × × ×2 × × ×2 × × ×2 × × ×

q

a b c d

1 3 2 21 1 1 00 1 2.5 1.54 0 2 02 2 1.5 1× 2 1 21 2 2 1.51 0 2 22 3 2 2× 1 2 20 1 2 0× 3 0 01 3 3 01 0 0 20 2 2.5 20 2 × 1× 0 1 0× 2 2 ×0 3 3 ×× 2 1.5 1.5× 3 2 0× 0 2.5 21 2 2 0.5× 0 1.5 1.51 2 × 12 1 × 02 2 0 0× 3 2 02 × 0 0

2 2 × ×× 2 3 12 × 0 11 1 × ×× 3 3 ×

2 2 0 ×0 × × 0× 0 2 11 × × 0.52 2 × ×× 2 2 ×1 × × ×× 2 2 0.50 2 × 2× 2 × 0× 0 2 0× 1 0 ×× 0 2 1× 2 3 0× 2 2 ×× 1 × ×× 0 2 1× 2 0 ×1 × × ×0 × × ×1 × × ×0 0 2 0× 0 2 0× 0 3 ×0 3 × ×2 × 0 ×× 0 2 0× 2 1.5 ×0 0 2 ×0 0 2 0× 3 × 00 0 × 1.5× 3 × ×2 × × ×3 × × ×2 × × ×2 × × ×2 × × ×2 × × ×2 × × ×

constancy (%)

a b c d

80.0 61.5 67.7 36.460.0 84.6 90.3 9.120.0 76.9 61.3 72.760.0 92.3 41.9 18.260.0 38.5 74.2 27.3

– 69.2 74.2 54.540.0 46.2 74.2 36.480.0 15.4 41.9 45.460.0 38.5 35.5 45.4

– 84.6 48.4 45.420.0 76.9 67.7 9.1

– 61.5 100.0 9.140.0 69.2 51.6 9.180.0 23.1 19.4 45.420.0 30.8 71.0 45.420.0 53.8 0 90.9

– 76.9 77.4 9.1– 100.0 61.3 –

20.0 69.2 71.0 –– 38.5 77.4 36.4– 69.2 64.5 9.1– 92.3 35.5 36.4

40.0 38.5 51.6 27.3– 84.6 32.3 36.4

60.0 30.8 – 45.440.0 84.6 – 9.180.0 38.5 3.2 9.1

– 69.2 38.7 18.2100.0 – 6.4 18.2

60.0 61.5 – –– 38.5 54.8 27.3

60.0 – 22.6 36.440.0 76.9 – –

– 53.8 61.3 –

60.0 30.8 19.4 –100.0 – – 9.1

0 23.1 58.1 27.380.0 – – 27.360.0 46.2 – –

0 69.2 35.5 –100.0 – – –

– 30.8 41.9 27.320.0 30.8 – 45.4

– 76.9 – 18.2– 23.1 61.3 9.1– 76.9 16.1 –– 15.4 32.3 45.4– 38.5 38.7 9.1– 30.8 54.8 –– 84.6 – –– 7.7 38.7 36.4– 1.5 19.4 –

80.0 – – –80.0 – – –80.0 – – –20.0 7.7 41.9 9.1

– 15.4 41.9 18.2– 23.1 51.6 –

20.0 53.8 – –60.0 – 12.9 –

– 15.4 48.4 9.1– 38.5 32.3 –

20.0 7.69 38.7 –20.0 7.69 29 9.1

– 46.2 – 18.220.0 7.7 – 36.4

– 61.5 – –60.0 – – –60.0 – – –60.0 – – –60.0 – – –60.0 – – –60.0 – – –60.0 – – –

Tab. 2. Continuation

Xmin – minimum, the lowest value of dataset,Xmax – maximum, the highest value of dataset,Q1 – first quartile,Q3 – third quartile,R – range,q – interquartile range.

Relevé sources of Table 2:a – community Sesleria-Pinus cembra, 5 relevés: Kanka (2008: tab. 11),b – asociation Seslerio variae-Piceetum Fajmonová 1978, 13 relevés: Fajmonová (1978: tab. 1)c – asociation Cortuso-Piceetum Fajmonová 1978, 31 relevés: Fajmonová (1985: p. 46 below), Fajmonová (1986: tab. 1,

relevés no. 1–25), Kubíček et al. (1996: p. 90), Kanka (2008: tab. 17, relevés no. 20 (2), 39 (6), 38 (5), 190 (17).d – asociation Polysticho lonchitidis-Piceetum W. Matuszkiewicz ex J. Matuszkiewicz 1977, 11 relevés: Kliment et al. (1982,

forest type 7405), Unar et al. (1984, tab. 37, relevé no. 2), Uhlířová et Bernátová (1986, relevé no. 1), Kanka (2008, tab. 9, relevés no. 78, 111, 121, 128, 139).

Acknowledgements

The author would like to thank to Ján Topercer (Comenius University in Bratislava, BotanicalGarden) for support and help with literature sources, and to Ladislav Jedlička (Comenius University

70

Poa stiriacaCorallorrhiza trifidaCampanula serrataCalamagrostis villosaSaxifraga paniculataOrthilia secundaAquilegia vulgarisRanunculus lanuginosusAlchemilla sp.Bistorta majorPedicularis verticillataBistorta viviparaMyosotis alpestrisSaxifraga wahlenbergiiCarex firmaErysimum witmanniiSelaginella selaginoidesCerastium *glandulosumDryas octopetalaHelianthemumgrandiflorumAstragalus norvegicusGymnocarpiumrobertianumFestuca tatraeLycopodium annotinumEuphorbia amygdaloidesMelica nutans

E0

Dicranum scopariumHylocomium splendensMnium spinosumPleurozium schreberiPlagiothecium curvifoliumTortella tortuosaCtenidium molluscumRhytidiadelphus triquetrusPolytrichum formosumRhizomnium punctatumPlagiothecium undulatumPlagiochila asplenioidesBazzania trilobataSphagnum capillifolium

Xmin

a b c d

× 0 0 00 0 × ×× × 0 0× × 0 00 × × 0× 0 × 0× 0 0 0× × 0 00 × 0 ×0 × × ×0 × × ×0 × × ×0 × × ×0 × × ×0 × × ×0 × × ×0 × × ×0 × × ×0 × × ×

0 × × ×0 × × ×

× 0 × 0× 0 × ×× × × 0× × 0 ×× 0 × ×

4 2 0 00 0 0 00 0 0 00 0 × 00 0 0 ×× 0 0 0× 0 0 00 0 0 00 0 × 00 × 0 00 × × 0× 0 0 ×0 × × 00 × × ×

Xmax

a b c d

× 4 3 21 2 × ×× × 3 2× × 3 22 × × 2× 2 × 1× 3 3 ×× × 3 31 × 1 ×3 × × ×3 × × ×2 × × ×2 × × ×2 × × ×2 × × ×1 × × ×1 × × ×1 × × ×2 × × ×

1 × × ×1 × × ×

× 2 × 2× 2 × ×× × × 4× × 3 ×× 3 × ×

4 4 4 44 3 4 35 5 3 45 3 × 34 3 3 ×× 3 3 2× 3 3 24 3 4 33 2 × 44 × 4 34 × × 3× 3 4 ×4 × × 23 × × ×

median

a b c d

× 0 0 00 0 × ×× × 0 0× × 0 00 × × 0× 0 × 0× 0 0 ×× × 0 00 × 0 ×0 × × ×0 × × ×0 × × ×0 × × ×0 × × ×0 × × ×0 × × ×0 × × ×0 × × ×0 × × ×

0 × × ×0 × × ×

× 0 × 0× 0 × ×× × × 0× × 0 ×× 0 × ×

4 3 0 32 2 0 20 3 0 03 2 × 00 2 0 ×× 2 0 0× 3 0 00 0 0 01 0 × 00 × 0 00 × × 0× 0 0 ×0 × × 00 × × ×

mode

a b c d

× 0 0 00 0 × ×× × 0 0× × 0 00 × × 0× 0 × 0× 0 0 ×× × 0 00 × 0 ×0 × × ×0 × × ×0 × × ×0 × × ×0 × × ×0 × × ×0 × × ×0 × × ×0 × × ×0 × × ×

0 × × ×0 × × ×

× 0 × 0× 0 × ×× × × 0× × 0 ×× 0 × ×

4 3 0 42 0 0 00 3 0 03 0 × 00 3 0 ×× 0 0 0× 3 0 00 0 0 00 0 × 00 × 0 00 × × 0× 0 0 ×0 × × 00 × × ×

Q1

a b c d

× 0 0 00 0 × ×× × 0 0× × 0 00 × × 0× 0 × 0× 0 0 ×× × 0 00 × 0 ×0 × × ×0 × × ×0 × × ×0 × × ×0 × × ×0 × × ×0 × × ×0 × × ×0 × × ×0 × × ×

0 × × ×0 × × ×

× 0 × 0× 0 × ×× × × 0× × 0 ×× 0 × ×

4 3 0 12 0 0 00 3 0 03 0 × 00 0 0 ×× 0 0 0× 2 0 00 0 0 00 0 × 00 × 0 00 × × 0× 0 0 ×0 × × 00 × × ×

Q3

a b c d

× 0 1 0× × 2 0× × 0 1.51 × × 0× 2 × 0× 2 0 ×× × 2 01 × 0 ×1 × × ×2 × × ×2 × × ×2 × × ×1 × × ×1 × × ×1 × × ×1 × × ×1 × × ×2 × × ×1 × × ×

1 × × ×1 × × ×

× 2 × 0× 2 × ×× × × 2× × 2 ×× 2 × ×

4 4 2 4 3 3 0 33 3 1 04 3 × 03 3 2 ×× 3 2 0× 3 2 02 2 2 03 0 × 12 × 0 13 × × 1.5× 2 0 ×4 × × 01 × × ×

R

a b c d

× 4 3 21 2 × ×× × 3 2× × 3 22 0 × ×× 2 × 1× 3 3 ×× × 3 31 × 1 ×3 × × ×3 × × ×2 × × ×2 × × ×2 × × ×2 × × ×1 × × ×1 × × ×1 × × ×2 × × ×

1 × × ×1 × × ×

× 2 × 2× 2 × ×× × × 4× × 3 ×× 3 × ×

0 2 4 44 3 4 35 5 3 45 3 × 34 3 3 ×× 3 3 2× 3 3 24 3 4 33 2 × 44 × 4 34 × × 3× 3 4 ×4 × × 23 × × ×

q

a b c d

× 0 1 00 1 × ×× × 2 0× × 0 1.51 × × 0× 2 × 0× 2 0 ×× × 2 01 × 0 ×1 × × ×2 × × ×2 × × ×2 × × ×1 × × ×1 × × ×1 × × ×1 × × ×1 × × ×2 × × ×

1 × × ×1 × × ×

× 2 × 0× 2 × ×× × × 2× × 2 ×× 2 × ×

0 1 2 31 3 0 33 0 1 01 3 × 03 3 2 ×× 3 2 0× 1 2 02 2 2 03 0 × 12 × 0 13 × × 1.5× 2 0 ×4 × × 01 × × ×

constancy (%)

a b c d

– 23.1 25.8 9.120.0 30.8 – –

– – 32.3 18.2– – 12.9 36.4

40.0 – – 9.1– 30.8 – 18.2– 38.5 9.7 –– – 35.5 9.1

40.0 – 3.2 –40.0 – – –40.0 – – –40.0 – – –40.0 – – –40.0 – – –40.0 – – –40.0 – – –40.0 – – –40.0 – – –40.0 – – –

40.0 – – –40.0 – – –

– 30.8 – 9.1– 38.5 – –– – – 36.4– – 32.3 –– 30.8 – –

100.0 100.0 38.7 72.780.0 61.5 16.1 54.540.0 92.3 25.8 18.280.0 53.8 – 9.140.0 61.5 32.3 –

– 61.5 41.9 18.2– 76.9 29.0 9.1

40.0 30.8 32.3 9.160.0 7.7 – 27.340.0 – 22.6 27.340.0 – – 27.3

– 38.5 19.4 –40.0 – – 9.140.0 – – –

in Bratislava, Faculty of Natural Sciences) for his lectures of biostatistics in time of author’s universitystudies. Moreover, to unlisted reviewer for his valuable comments on first version of the manuscript.This study was supported by Slovak grant agency VEGA, grant No. 1/2347/05 and recent grant No. 2/0059/11.

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Appendix 1: The original table form of Tüxen, Ellenberg (1937: 173, Tabelle 1) with the original comment of theauthors

3) Die Festsetzung dieser Zählenwerte ist naturgemäß willkürlich, da durch + und 1 lediglich verschiedenen großeIndividuenzahl bei gleichen Deckungsgrand (unter 5 %) bezeichnet wird. Wir haben uns für die Werte 2,5 und 0,1 aufGrund freundlichst zur Verfügung gestellten Berechnung von Herrn Forsting. “Meijer Drees entschieden.” (Tüxen,Ellenberg 1937: 173).

Appendix 2: Small phytosociological English-Slovak statistical vocabularyEnglish terms are according to Zar (1996); those in brackets according to Sokal, Rohlf (1997); Slovak terms are usedaccording to cited Slovak biostatistical literature.For purposes of syntaxonomy, one phytosociological relevé is an individual observation, the smallest sampling unit (cf. Sokal,Rohlf 1997: 8). Set of relevés represents a sample of observations (cf. Sokal, Rohlf 1997), which substitutes in a (biostatistical) study a population (statistical use of the term, not the biological one). The relevé is a basic object ofobservation, and variables are its species and other characteristics (elevation, orientation and inclination of slope, etc.). As variates could be designated species’ cover-abundance values. A parameter describes or characterizes a population; a statistic is an estimation of a population parameter from the sample (within the sample) (Zar 1996).interquartile range/semi-quartile range – kvartilové rozpätiemeasures of central tendency (statistics of location) – charakteristiky polohymeasures of dispersion (statistics of dispersion) – charakteristiky variabilityordinal scale – poradová (ordinálna) stupnicaparameters – parametre: veličiny charakterizujúce vlastnosti základného súborupopulation – základný súbor (ide o štatistické, nie biologické použitie termínu population)quartile deviation/semi-interquartile range – kvartilová odchýlkarandom sampling – náhodný výber (prevažnú časť fytocenologických zápisov nemožno považovať za náhodný výber; ten sariadi určenými princípmi – pozri napr.: Jedlička, 1984, p. 18; Lepš, 1996, p. 6–7).range – variačné rozpätieratio – pomer, v štatistike druh odvodených znakov z primárnych znakov objektovratio scale – pomerová stupnicasample – štatistický výber (výber), časť základného súborustatistics (sing. statistic) – výberové charakteristiky: veličiny, ktoré v sebe koncentrujú informácie o výbere a vypovedajú o jeho vlastnostiachstatistics (descriptive statistics) – opisné (deskriptívne) štatistické charakteristikyvariable (or variate: Zar 1996: 2) – znak (t.j. označenie vlastnosti “characteristic/character” biologického objektu)variate (Sokal, Rohlf 1997: 12; see note at p. 13) – variant (daná hodnota znaku)

73

Mengenskala nach Braun-Blanquet

5

4

3

2

1

+

Interval desDeckungsgrades(% der Aufnahmefläche)

75–100

50–75

25–50

5–25

} 0–5

Mittel

87,5

62,5

37,5

15

2,53)

0,13)

Abstrakt

Použitie aritmetického priemeru je podľa štatistickej metodiky obmedzené na dáta v pomerovej a intervalovej stupnici. Vo fytocenológii sa napriek tomu používa najčastejšie na hodnotenie priemernejpokryvnosti druhu vypočítavanej z Braun-Blanquetovej stupnice početnosti a pokryvnosti, ktorá jeordinálneho rázu. Namiesto aritmetického priemeru sa navrhuje použitie vhodnejších deskriptívnych(opisných) štatistík. Článok zhŕňa základné skutočnosti o štatistických vlastnostiach Braun--Blanquetovej stupnice početnosti a pokryvnosti a prislúchajúcich opisných štatistikách. Ich použitie jepredvedené na príklade štyroch spoločenstiev radu Athyrio filicis-feminae-Piceetalia Hadač ex Hadačet al. 1969.

Peter Kučera: Použitie deskriptívnych štatistických charakteristík vo fytocenológii

74

ACTA BOTANICA UNIVERSITATIS COMENIANAE

Volume 46

Vydala Univerzita Komenského v Bratislave vo Vydavateľstve UK

Technická redaktorka: Alena Zvěřinová

ISBN 978-80-223-3113-5ISSN 0524-2371