Research Article ii FF o o r r e e s s t t doi: 10.3832/ifor1370-008 Biogeosciences and Forestry vol. 9, pp. 330-336

Ectomycorrhizal fungal community associated with autochthonous white poplar from

Marina Katanić (1), Tine We analyzed the community of ectomycorrhizal fungi of an autochthonous Grebenc (2), Saša Orlović (1), white poplar (Populus alba L.) stand in the Kovilj- marshes (Ser- Milan Matavuly (3), Branislav bia), and examined its seasonal dynamics. Ectomycorrhizal types were identi- (1) (2) fied by combining morphological and anatomical descriptions with molecular Kovačević , Marko Bajc , methods (sequencing of ITS region of ribosomal DNA). In two seasons, 20 ecto- (2) Hojka Kraigher mycorrhizal types were recorded, from which 11 types were identified to the species level, six were determined to the genus level, two types were deter- mined to the family level and one type remained unidentified. Number of ectomycorrhizal types, number of fine roots, percentage of vital mycorrhizal roots, diversity indexes and abundance of exploration types did not differ sig- nificantly between autumn and spring. During both seasons, the most abun- dant types were: Entoloma sp., Tuber maculatum, Cenococcum geophilum, Tuber rufum and Peziza sp. Due to the high variation of the ectomycorrhizal types-based Shannon-Weaver diversity index in poplar stands, and the fact that poplars form dual mycorrhizal association, this index is not recommended as a reliable index for bioindication in poplar.

Keywords: Ectomycorrhiza, Populus alba, Diversity, Nature Reserve, Seasonal Dynamics, Morphological-Anatomical Characterization, Molecular Identification

Introduction has rarely attracted the interest of resear- improvement of soil structure by the extra- Poplars are world-wide distributed tree chers, several prominent ECM species are matrical hyphal network (Molina et al. species that combine commercial impor- thoroughly studied in poplars, in particular 1992, Smith & Read 2008, Quoreshi 2008). tance with biotechnological advantages, in white poplar (Populus alba L.), including The functional compatibility and stress to- such as rapid growth, simple in vitro propa- the most expensive white truffle (Tuber lerance of ectomycorrhizal types is species gation and availability of genetic transfor- magnatum Pico - Angelini & Granetti 1995). specific and depend on both partners. The- mation systems (Klopfenstein et al. 1997, Seedlings colonized with compatible ECM refore, information on the ECM community Kaldorf et al. 2004). They are used in agro- fungal species and strains are favored in structure can provide valuable information forestry systems (Eichhorn et al. 2006), making contacts with water and nutrients, about physiology of forest trees and func- short rotation forestry (Klašnja et al. 2006) as well as with other organisms in the soil tioning of forest ecosystems (Kraigher et and phytoremediation, as they cycle large (Kraigher 1996). In addition to the increa- al. 2007). amounts of water and grow rapidly by pro- sed nutrient uptake, mycorrhizas offer nu- Limited data on ECM community in white ducing large amount of biomass, due to merous other benefits to symbiontic tree poplar from natural stands are available their deep root systems (Newman et al. species, such as enhanced plant efficiency and its seasonal dynamics remain unclear 1997). in absorbing water, reduced fertilization (Jakucs 2002). The aim of this work was to Poplars in plantations and natural stands and irrigation requirements, increased investigate the ECM community in a case regularly form dual associations with ecto- drought tolerance, increased pathogen re- study location represented by an autoch- mycorrhizal (ECM) and arbuscular mycor- sistance, protection against damage from thonous white poplar stand in the Special rhizal (AM) fungi (Karlinski et al. 2010) that heavy metals and other pollutants, mitiga- Nature Reserve “Kovilj-Petrovaradin mar- are known to prefer different soil depths tion of various plant stresses, improve- shes” in Serbia and to analyze seasonal (Neville et al. 2002). While AM community ment of seedling growth and survival, and changes in the community between spring and autumn.

(1) University of , Institute of Lowland Forestry and Environment, Antona Cehova Materials and methods 13, 21000 Novi Sad (Serbia); (2) Slovenian Forestry Institute, Večna pot 2, 1000 Ljubljana (Slovenia); (3) European University, Faculty of Pharmacy, Trg mladenaca 5, 21000 Novi Sad Description of site and sampling (Serbia) ECM roots were isolated from soil sam- ples collected in the Special Nature Re- ′ @ Marina Katanić ([email protected]) serve, close to Novi Sad, Serbia (45° 12 N, 19° 58′ E, elevation 78 m a.s.l.). According Received: Jun 04, 2014 - Accepted: Jul 10, 2015 to the results from a nearby measuring sta- tion (Rimski Šančevi), the average annual Citation: Katanić M, Grebenc T, Orlović S, Matavuly M, Kovačević B, Bajc M, Kraigher H precipitation (1951-2010) was 625 mm and (2015). Ectomycorrhizal fungal community associated with autochthonous white poplar from the average yearly temperature was 11.4 °C Serbia. iForest 9: 330-336. – doi: 10.3832/ifor1370-008 [online 2015-11-12] (Stanojević 2012). Climate is a Dfa subtype of temperate continental climate with July Communicated by: Paola Mairota and August as the warmest months (Re- public Hydrometeorological Service of Ser-

© SISEF http://www.sisef.it/iforest/ 330 iForest 9: 330-336 Katanić M et al. - iForest 9: 330-336

y n r bia - http://www.hidmet.gov.rs/). The sam- criptions published in Agerer (2008), Age- and i is the number of mycorrhizal tips t

s pling site was occasionally flooded and the rer et al. (2006), and Agerer & Rambold of individual ECM type; e e H S H r soil type was classified as fluvisol, loamy (2014). Based on the presence and abun- (iii) Evenness ( ) = / log , where is the o form (Katanić 2014). The sampling was per- dance of emanating elements, ECM types Shannon-Weaver’s diversity index and S F formed in a naturally grown autochtho- were also classified into the exploration ty- is the number of ECM types; d n nous white poplar (Populus alba L.) stand pes proposed by Agerer (2001). (iv) Equitability (J) = H/Hmax, where H is the a -1 of 50-60 years old trees (100 trees ha ) Molecular identification was based on Shannon-Weaver’s diversity index and s e mixed with scarcely abundant Acer negun- nucleotide sequencing of ITS regions (In- Hmax is the theoretical maximum H assu- c

n do L. and Robinia pseudoacacia L. ternal Transcribed Spacer) in nuclear ribo- ming that each ECM type was equally e i Five mature white poplar trees were ran- somal DNA. This is considered the best mo- abundant; c

s domly selected. Two soil samples per tree lecular marker for fungi identification (Kõl- (v) Berger-Parker’s evenness index (BP) = 1 o were taken in autumn (September 2009) jalg et al. 2013). After DNA extraction from - (Nmax/N), where Nmax is the number of e ® g and spring (March 2010) at a distance of 5-20 root tips with a PlantDNeasy Mini Kit mycorrhizal tips of the most frequent o i about 1 m from the tree trunk. In total, ten (Qiagen, Hilden, Germany) from each ECM ECM type and N is the number of all my- B soil samples were collected in each season. type, the ITS region was amplified using corrhizal tips. – A soil corer of 274 ml volume and reaching the ITS 1f and ITS 4 primer pair (Gardes & Data of two soil core samples were joint t s 18 cm depth was used for collecting stan- Bruns 1993). DNA fragments were sepa- and single tree was used as a statistical e r dardized soil core samples (Kraigher 1999). rated in and excised from agarose gel and unit. The Student t-test was used to test o ®

F Soil core samples were stored at 4 °C for up purified with Wizard SV Gel and PCR the significance of differences in the num- i to one month. One day prior to analysis, Clean-up System® (Promega Corporation, ber of ECM types, vital ECM root tips, old, soil samples were submerged in water and Madison, WI, USA). Sequencing was per- non-turgescent and non-mycorrhizal roots, all fine roots were carefully washed from formed commercially at Macrogen Inc. total fine roots, percentage of vital root soil. Vital ECM root tips were separated (Seoul, Rep. of Korea). Species, genus or tips and abundance of exploration types from old, non-turgescent and non-mycor- family of ECM fungi were determined by between autumn and spring. In order to fit rhizal (ONN) root tips in water under a dis- comparing with the sequences deposited the normal distribution, data were trans- secting microscope. All fine root tip cate- in the GenBank (http://www.ncbi.nlm.ni formed as follows: count data were trans- gories, including all identified types of h.gov/genbank/index.html) and Unite (Aba- formed according to square root transfor- ECM, were counted as total number in indi- renkov et al. 2010) databases. mation (Bartlett 1936), while percentage vidual soil core sample. values were transformed according to arc- Data analysis sine transformation using the Bliss formula Identification of ectomycorrhizae Diversity indexes were calculated per (Snedecor & Cochran 1976). The non-trans- ECM types were identified by combining sample and per site (i.e., by pooling the formed data are presented in Tabs. 2 and 3. morphological and anatomical approach ECM community data) following the formu- The Mann-Whitney U test was used to test with molecular methods. Morphological las given by Atlas & Bartha (1981) and Tay- the significance of differences in diversity and anatomical characteristics of each ECM lor et al. (2000): indexes. All statistical analyses were per- ® root type were assessed by a binocular (i) Species richness (d) = (S - 1) / log10 N, formed using the package STATISTICA ver- Olympus SZX 12 (light source: Olympus where S is the number of ECM types and sion 12 (StatSoft Inc., Tulsa, OK, USA). Highlight 3100, daylight filter) and DIC N is the number of all mycorrhizal tips; (Nomarski) microscope Olympus BX 51 (ii) Shannon-Weaver’s diversity index (H) = Results (magnification 100-2000×) following Age- C/N (N log N – Σ ni log ni), where C = 2.3, Overall, twenty ECM types were determi- rer (1991), Kraigher (1996), and ECM des- N is the number of all mycorrhizal tips ned at the examined site in the two investi- gated seasons. Eleven types were identi- fied to the species level, six to the genus Tab. 1 - Identification of ectomycorrhizal (ECM) types from a mature white poplar level, while other three were determined (Populus alba L.) stand at the Special Nature Reserve Kovilj-Petrovaradin marshes to the family level or remained unidentified (Serbia). Identifications are based on morphological-anatomical characters and nrITS (Unknown type KR - Tab. 1). Cenococcum DNA sequence comparison with the international nucleotide sequences databases. geophilum, Entoloma sp., Genea verrucosa, GenBank accession numbers are given for the analyzed ECM types. Exploration types Inocybe maculata, Inocybe sp., Peziza sp., were assessed after Agerer (2001). Sebacina incrustans, Tomentella sp. 2, Corti- nariaceae sp., Tuber rufum and Tuber macu- Ectomycorrhizal type Nucleotide database Exploration type latum were recorded in both seasons. He- accession number beloma vaccinum, Inocybe cf. rimosa, Ino- C. geophilum HG937623 short distance cybe obsoleta, Thelephoraceae sp., and To- Genea verrucosa HG937624 short distance mentella sp. 1 were observed only in spring, Hebeloma vaccinum HG937625 medium distance fringe subtype while Clavulina sp., Hymenogaster tener, Hymenogaster tener HG937626 short distance Inocybe cf. meliolens and Unknown type KR Inocybe cf. meliolens HG937629 short distance were recorded only in autumn (Fig. 1a, Fig. Inocybe cf. rimosa HG937627 short distance 1b). Inocybe maculata HG937628 short distance No significant differences between sea- Inocybe obsoleta HG937630 short distance sons were found as for the number of ECM Sebacina incrustans HG937631 short distance types, vital ECM roots, old, non-turgescent Tuber maculatum HG937633 short distance Tuber rufum HG937632 short distance and non-mycorrhizal roots, total number of Clavulina sp. HG937634 short distance fine roots and percentage of vital ECM root Cortinariaceae sp. HG937640 short distance tips. However, all values were higher in Entoloma sp. HG937635 medium distance smooth subtype spring (Tab. 2). Inocybe sp. HG937636 short distance There were no significant differences bet- Peziza sp. HG937637 contact ween seasons in species richness index, Tomentella sp. 1 HG937638 short distance Shannon-Weaver’s index, Evenness, Equi- Tomentella sp. 2 HG937639 short distance tability and Berger-Parker’s index (Tab. 2). Thelephoraceae sp. HG937641 short distance Diversity indexes revealed that number of Unknown type KR - short distance species, relative abundance of individual

331 iForest 9: 330-336 Ectomycorrhizal fungal community in autochthonous white poplar y r

Fig. 1 - Ectomycorrhizal community structure t s

in a mature white poplar stand in Serbia, e based on sampling of 10 soil samples in spring r o F

(a) and 10 soil samples in autumn (b). d n a

s e c n e i c s o e g o i B

–

t s e r o F i

Fig. 2 - Relative abundance of ectomycorrhizal fungal families in a mature white poplar stand in Serbia, based on sampling of 10 soil samples in spring (a) and 10 soil samples in autumn (b)

iForest 9: 330-336 332 Katanić M et al. - iForest 9: 330-336 y r

t Tab. 2 - Comparison of total and average values (± standard erorr) of number of ectomycorrhizal (ECM) types, vital ECM root tips, s

e old, non-turgescent and non-mycorrhizal roots (ONN), total fine roots and diversity indexes from a mature white stand located in r Serbia between spring and autumn. (a): p-values after the t-test; (b): p-values after the Mann-Whitney’s U test. o F

d Spring Autumn n a Parameter p-value Total value Average value Total value Average value s per site per tree per site per tree e c Number of ECM types 16 5.10 ± 0.60 15 4.90 ± 0.20 0.838 a n a e Number of vital ECM root tips 4300 430 ± 133.6 3030 303 ± 65.5 0.511 i a c Number of ONN root tips 22140 2214 ± 511.6 19335 1933.5 ± 148.2 0.686 s Total number of fine root tips 26440 2644 ± 607.7 22365 2236.5 ± 141.9 0.609 a o a e % of vital ECM Root tips 16 16.20 ± 2.90 14 13.90 ± 3.10 0.569 g Species richness index 4.13 1.62 ± 0.20 4.02 1.63 ± 0.08 0.754 b o i Shannon-Weaver index 1.95 1.01 ± 0.12 2.00 1.15 ± 0.06 0.403 b B Evenness 0.70 0.626 ± 0.06 0.74 0.723 ± 0.03 0.174 b –

b

t Equitability 1.62 1.44 ± 0.13 1.70 1.67 ± 0.07 0.174 s Berger-Parker index 0.58 0.38 ± 0.04 0.57 0.48 ± 0.04 0.117 b e r o F i species, equitability and evenness between The most abundant ECM fungal family higher than those colonized by Ascomy- species, and dominance rate of the most was Entolomataceae, followed by Tubera- cota fungi (in spring 68.5 vs. 31.5 %, respec- abundant species did not differ signifi- ceae and Cortinariaceae. In both seasons tively, in autumn 62 vs. 38%, respectively - cantly between seasons, i.e., the ECM com- the three above-mentioned families repre- data not shown). munity structure did not change across sented 70% of the total number of all ECM The most abundant exploration type (ET) seasons. root tips (Fig. 2a, Fig. 2b). was short-distance ET followed by medium In both seasons, the same five ECM types In terms of species richness, the family -distance ET and contact ET. Long-distance dominated and represented about 80% of Cortinariaceae was the richest, with six ET was not observed. Abundance of each the total number of ECM root tips. The species in spring and four in autumn (Fig. ET did not significantly differ between the most abundant ECM type was Entoloma sp. 3). The number of Basidiomycota ECM sym- seasons (Tab. 3). with over 40%, followed by Tuber macula- bionts was higher than those of Ascomy- tum, Cenococcum geophilum, Tuber rufum cota, both in spring (11 vs. 5, respectively) Discussion and Peziza sp. Relative abundance of these and in autumn (10 vs. 5, respectively). Simi- The community of ectomycorrhizal fungi ECM types was similar in both seasons (Fig. larly, the contribution of root tips colo- in this case study represented by a mature 1a, Fig. 1b). nized by Basidiomycota ECM fungi was autochthonous white poplar stand located in the Special Nature Reserve “Kovilj-Petro- varadin marshes” (Serbia) is one of the few studies (Jakucs 2002, Katanić et al. 2010, Katanić 2014) on ECM diversity in white poplar and, up to our knowledge, the only one in a mature natural stand. The average amount of vital ECM root tips (1106 dm-3 in autumn and 1570 dm-3 in spring) was considerably lower than in ma- ture stands of other species such as beech (1460-15 800 dm-3 - Mašek & Grebenc 2011) and spruce (4309-6716 dm-3 - Kraigher 1999). The number of all fine roots per soil volume in poplar stands was even more va- riable. In this study, such value was 9651 dm-3 in spring and 8163 dm-3 in autumn, while ranged from 2599 dm-3 in control plants irrigated with water to 4573 dm-3 in plants treated with the anti-ozonant ethy- lenediurea in an ozone-sensitive clone (Ka- tanić et al. 2014). Krpata et al. (2008) counted 17 350-42 630 dm-3 ECM root tips at a site with aspen contaminated by heavy metals. The relatively low number of ECM Fig. 3 - Number of ectomycorrhizal fungal taxa within fungal families recorded in roots in our white poplar stand reflects the mature white poplar stand located in Serbia in spring and autumn. regular flooding occurring in the studied area and the dual colonization with ECM and AM fungi. In the same poplar trees, Tab. 3 - Relative abundance (± standard error, %) of ectomycorrhiza exploration types Katanić et al. (2013) recorded values of in a mature white poplar stand in Serbia in spring and autumn, and significance of Stu- root length colonization with ECM and AM dent t-test for the effect of season. fungi of 16.7 and 8.8 %, respectively. The 20 ECM types recorded in total are a Exploration type Spring Autumn t-test (p-value) considerably lower number compared to Contact type 1.87 ± 1.01 2.59 ± 2.59 0.754 some previous studies. On individual Popu- Short distance 65.23 ± 9.38 60.68 ± 8.42 0.648 lus tremula trees in an old-growth mixed Medium distance 32.90 ± 9.09 36.72 ± 9.09 0.724 forest, Bahram et al. (2011) found 122 ECM Long distance 0 0 - fungal species. Analyzing ECM community

333 iForest 9: 330-336 Ectomycorrhizal fungal community in autochthonous white poplar y

at two sites with 30-40 m tall white poplars ECM types (in spring and autumn, respec- dance with Courty et al. (2008) who re- r t

during three years, Jakucs (2002) recorded tively) contributed to the rest. This finding corded the highest number of vital ECM s e

70 ECM types, while Krpata et al. (2008) is in accordance with the evidence that the roots in April, while the lowest value was r found 54 ECM types at a heavy metal pol- ECM community varied according to a loga- observed in September. The absence of o F luted site with 20 to 25-year-old aspen rithmic distribution of abundances (Dahl- significant differences between autumn d trees. However, results of our research are berg 2001, Pickles et al. 2010). and spring in the number of ECM types and n a in accordance with Visser et al. (1998), who In the Kovilj-Petrovaradin marshes, the diversity indexes in this study is in accor- s recorded 22 ECM types in a mixed forest genus Inocybe was the most abundant one, dance with the results of De Roman & De e c

dominated by American aspen, and Kaldorf with five ECM types, while genera Tuber Miguel (2005). In both seasons the same n e et al. (2004), who observed 23 morphoty- and Tomentella had two members. Analy- three fungal families were the most abun- i c

pes in a five-year-old experimental aspen zing the diversity of mycorrhizal fungi in dant (Entolomataceae, Tuberaceae and s plantation. In addition, at three experimen- mixed deciduous stand, Lang et al. (2011) Cortinariaceae). In contrast, Richard et al. o e tal sites with poplars Karlinski et al. (2013) noticed that Tomentella and Inocybe were (2011) found that the relative abundance of g o found in total 27 ECM fungal taxa. In a the most abundant and contributed mostly two out of three the most abundant fami- i B white poplar plantation, Katanić et al. to the species richness. In the ECM commu- lies (Russulaceae and Cortinariaceae) sho- – (2010) preliminary recorded 15 ECM types. nity associated with aspen grown at the wed significant seasonal shifts. t

However, when ECM diversity was studied site contaminated with heavy metals, the In both seasons, Basidiomycota domina- s e seasonally, a total of 30 ECM types were most abundant taxonomic groups were ted the examined ECM community, confir- r o

observed (Katanić 2014). It seems that the Tomentella (17), Inocybe (6), Cortinarius (5), ming numerous studies on poplars (Cripps F number of ECM fine roots and the diversity Hebeloma and Tuber with 3 operational ta- 2001, 2003, Krpata et al. 2008, Karlinski et i of ECM types in poplar are highly variable, xonomic units (Krpata et al. 2008). At three al. 2013, Katanić et al. 2014) where the and the naturalness of the studied Special sites with poplar clones, Karlinski et al. Basidiomycota group was the most abun- Nature Reserve does not contribute to in- (2013) recorded the dominance of Cortina- dant and had more members in compari- crease ECM diversity as compared with riaceae, Thelephoraceae and Tricholoma- son to Ascomycota. other poplars’ sites. taceae that constituted nearly 90% of the The most abundant exploration type in Values of Shannon-Weaver’s diversity in- mycorrhizal community. both seasons was short-distance ET and dex (1.95 in spring and 2.00 in autumn) are Inocybe species are well-known coloni- then medium-distance ET. Agerer (2001) in accordance with results obtained by De- zers of ECM plants on disturbed or pioneer found a relationship between ETs and their Bellis et al. (2006). These authors recorded sites (Nara et al. 2003). Investigating my- potential ecological roles. Jakucs (2002) a similar Shannon-Weaver diversity index corrhizal fungi associated with aspen at noticed that in two forests adapted to simi- of 2.00 in an aspen-dominated plot by three different sites, Cripps (2003) obser- lar ecological conditions, dominant ECM using morphological characterization of ved 54 ECM fungi and 14 species belonged types belonged to the same ET. Analyzing ECM types, but a considerably higher index to the genus Inocybe. Tomenteloid fungi ET of ECM community, Rudawska et al. was recorded (3.00) when molecular iden- are the most frequent and widespread (2011) concluded that abundance of a par- tification tools were applied. Values of the ECM partners of deciduous and evergreen ticular ET is related to the soil chemistry, Shannon-Weaver’s diversity index obtained tree species in the forests of Europe and since occurrence of contact ET was related in extreme conditions are controversial. On North America (Gardes & Bruns 1996, Dahl- to high nutrient content. On the other the one hand, an ozone sensitive poplar berg 2001). In the white poplar forest adap- hand, medium-distance fringe exploration clone showed lower values of Shannon- ted to dry conditions of the Hungarian type was abundant at a sites contaminated Weaver’s index, namely 1.60 in anti-ozo- plain, Tomentella group was a minority with heavy metals (Rudawska et al. 2011, nant protected plants and 1.21 in control component of the ECM community (Jakucs Karlinski et al. 2013). We assume that the plants (Katanić et al. 2014). On the other 2002). Similar observations were made in similar distribution of ETs observed at our hand, Krpata et al. (2008) recorded rela- the occasionally flooded Kovilj-Petrovara- site in spring and autumn results from simi- tively high value of Shannon-Weaver’s di- din marshes. Investigating the molecular lar ecological conditions. versity index (2.00) in aspen trees grown at diversity and the ecological specificity of a site contaminated with heavy metals. truffles originating from the mid-west of Acknowledgements Due to the high variation of the ECM-based Balkan Peninsula, Marjanović et al. (2010) The study was co-financed by the Slove- Shannon-Weaver’s diversity index for pop- recorded 12 species from this group, inclu- nian Research Agency through the Re- lar stands, and the fact that poplars form ding Tuber rufum and T. maculatum. Ceno- search Programme P4-0107 “Forest Biolo- dual mycorrhizal association, this index can coccum geophilum is a cosmopolitan ECM gy, Ecology and Technology”, through the not be considered reliable for bioindica- fungus with a wide range of hosts and Scholarship Ad futura (OMEGA D.O.O., for tion, as otherwise proven for spruce (Krai- habitats. It is a frequent and abundant ECM MK) and project III43007 “Studying climate gher 1999). None of the above-mentioned type adapted to environment under stress change and its influence on the environ- authors compared fine roots data, species (LoBuglio 1999) that was recorded in al- ment: impacts, adaptation and mitigation” abundance or diversity indexes of poplar most half of the samples in our study. Di financed by the Ministry of Education, Sci- stands among seasons. However, in our Pietro et al. (2007) noted that under ex- ence and Technological Development of study differences between seasons did not treme stress, ECM formed by Cenococcum the Republic of Serbia within the frame- show any significant effect on diversity of geophilum has the ability to survive better work of integrated and interdisciplinary the ECM community. than ECM of some other fungi, and that research. ECM community associated with white this species is particulary efficient in the poplars in the Kovilj-Petrovaradin marshes protection of fine roots from drought References consisted of few abundant and numerous stress. However, during the three-year sur- Abarenkov K, Nilsson RH, Larsson K-H, Alexan- infrequent ECM types, in accordance with vey of two ECM communities associated der IJ, Eberhardt U, Erland S, Høiland K, Kjøller previous research (De Roman & De Miguel with poplars in dry conditions, Jakucs R, Larsson E, Pennanen T, Sen R, Taylor AFS, 2005, O’Hanlon & Harrington 2012). Koide (2002) did not observe the presence of C. Tedersoo L, Ursing BM, Vrålstad T, Liimatainen et al. (2007) reported abundant ECM types geophilum, while in our study it was found K, Peintner U, Kõljalg U (2010). 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