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Citation: Shiryaev, A.G.; Zmitrovich,
I.V.; Senator, S.A.; Minogina, E.N.;
Tkachenko, O.B. How Poor Is
Aphyllophoroid Fungi Diversity in
the Boreal Urban Greenhouses of
Eastern Europe?. J. Fungi 2023,9,
1116. https://doi.org/10.3390/
jof9111116
Academic Editor: Lei Cai
Received: 19 October 2023
Revised: 10 November 2023
Accepted: 16 November 2023
Published: 17 November 2023
Copyright: © 2023 by the authors.
Licensee MDPI, Basel, Switzerland.
This article is an open access article
distributed under the terms and
conditions of the Creative Commons
Attribution (CC BY) license (https://
creativecommons.org/licenses/by/
4.0/).
Fungi
Journal of
Article
How Poor Is Aphyllophoroid Fungi Diversity in the Boreal
Urban Greenhouses of Eastern Europe?
Anton G. Shiryaev 1, * , Ivan V. Zmitrovich 2, Stepan A. Senator 3, Elena N. Minogina 4and Oleg B. Tkachenko 3
1Institute of Plant and Animal Ecology, Ural Branch of the Russian Academy of Sciences,
620144 Ekaterinburg, Russia
2V.L. Komarov Botanical Institute, Russian Academy of Sciences, 197376 Saint Petersburg, Russia;
iv.zmitrovich@gmail.com
3N.V. Tsitsin Main Botanical Garden, Russian Academy of Sciences, 127276 Moscow, Russia;
stsenator@yandex.ru (S.A.S.); ol-bor-tkach@yandex.ru (O.B.T.)
4Botanical Garden, Ural Branch of the Russian Academy of Sciences, 620144 Ekaterinburg, Russia;
minogina71@mail.ru
*Correspondence: anton.g.shiryaev@gmail.com
Abstract:
It is generally accepted that mycobiota diversity in urban greenhouses is poorer than in
natural ecosystems, but our knowledge on this field of research is fragmentary. Here, we present
the results of a long-term study of aphyllophoroid macrofungi (Basidiomycota) forming fruitbodies
on non-native sub/tropical woody and herbaceous plants in the greenhouses of Saint Petersburg,
Moscow, and Ekaterinburg botanical gardens located in the hemiboreal vegetation subzone of Eastern
Europe. Over 20 years of research, fruitbodies of 58 species of aphyllophoroid fungi have been
identified. Fungal species that developed on the wooden structures of greenhouses and building
materials made of local wood are discussed separately. The list of fungi on non-native substrates is
dominated by saprobes (93.1% of total list) as well as mycorrhizal with basidiomata on plants (8.6%).
Phytopathogens have the lowest number (7.0%), and
3
4
of species are widespread locally. Non-native
plants are dominated by native fungal species (78.9%), while the percentage of non-native species
is low (21.1%). In the three surveyed cities, the area of the studied greenhouses is 2.8 ha, and not a
single species of fungi has been found twice on the same substrate. Half of the identified species
are characterized by a single specimen (29 species/50.9%). Hymenochaete rheicolor was discovered
in Russia for the first time and its known distribution is discussed. Only six (Antrodia gossypium,
Hyphodontia arguta,Lyomyces sambuci,Peniophora cinerea,Ramariopsis kunzei, and Trechispora farinacea)
local species (10.5%) were collected in all the three cities. The
α
-diversity of mycobiota (mean number
of species per site, Shannon Index, and Menhinick Index) in the Ekaterinburg’s greenhouses is
1
.2–3.0 ti
mes lower compared to suburban forest parks and old-growth natural forests, while
β
-
diversity (Whittaker Index, Jaccard Index, and Morisita–Horn Index), on the contrary, is
2.1–7.7 time
s
higher. With the plants’ age, the probability of detecting fungi on them increases significantly. In
greenhouses, phytopathogenic aphyllophoroid macrofungi are collected on woody plants only, but
the probability of their development is not related to the plants’ age.
Keywords: botanical garden; non-native fungi; invasion; pathogen; plant disease
1. Introduction
The study of the diversity and dynamics in various components of urban ecosystems
is one of the most important areas of modern research in biology and ecology [
1
]. Forest
fragments that have survived the process of urbanization are a characteristic feature of
many large cities and are often represented by unique flora complexes and associated
mycobiota [
2
,
3
]. The vegetation of botanical gardens, which are an integral part of such
forests, is a reflection of urban nature—with potentially widespread non-native species,
which coexist with stable local relatives and purposefully introduced flora [4].
J. Fungi 2023,9, 1116. https://doi.org/10.3390/jof9111116 https://www.mdpi.com/journal/jof
J. Fungi 2023,9, 1116 2 of 23
With non-native plants, many species of non-native fungi are unintentionally spread
into urban settings and botanical gardens, which can cause significant environmental
and economic damage [
5
]. In plantings of non-native plants in botanical gardens, a great
number of non-native fungal species are concentrated. For some invasive fungal species,
plantings of botanical gardens are a refuge for acclimatization to new environments and
a springboard for introduction into the local natural forests [
3
,
6
]. Although the marked
impact of fungal introductions on ecosystems is recognized [
4
,
7
,
8
], general knowledge
of the geographic distribution and distribution patterns of non-native fungi is far from
complete [2,9,10], and needs to be constantly updated.
The maximum concentration of non-native plant species is characteristic of green-
houses, where the plants are sub/tropical along with their own number of trophically
associated exotic fungal species. Under favorable, but limited conditions, such fungi can
spread into local nature causing disease or death of local plants [
8
,
11
–
15
]. Nevertheless,
there are very little data on the fungal diversity in greenhouses.
Among various groups of macrofungi developing in the greenhouses of Europe, the
bulk of the data relate to a targeted study of the species composition of agaricoid fungi.
A number of greenhouses located in the nemoral regions of Central Europe (Austria,
Germany, Poland, and Czech Republic) were studied, in which the biodiversity of agaricoid
fungi was assessed based on the presence of fruitbodies in the soil [
6
,
7
,
10
,
14
,
16
–
19
]. It
has been demonstrated that the number of fungal species in greenhouses is significantly
less compared to natural conditions, and mainly local fungal species develop, and the
proportion of non-native sub/tropical fungi varies in the range of 10–20% of the total
number of species. Agaricoid fungi in greenhouses are humus saprobes that have been
sporadically found to be capable of exhibiting pathogenic or mycorrhizal activity [
19
,
20
].
The total number of agaricoid fungi in the greenhouses of these countries is 187 species.
Due to the small number of other groups of macrofungi in the greenhouses of Europe,
they are rarely the objects of special research. For example, in the greenhouses of the
Botanic Garden of the Institute of Botany in Graz (Austria), there are certain isolated data
on the findings of aphyllophoroid fungi on wooden structures and building materials, but
very few species of fungi have been collected on living or dead parts of non-native plants.
In the sub/tropical greenhouses, on the outside of a very moist flower pot with substrate
and the seedling of a date palm species, the poroid Rigidoporus lineatus (Pers.) Ryvarden
was collected [
7
]. It was also collected in the Budapest greenhouse. This is a common
polypore in the tropics, but rare in Europe [
12
,
15
]. Basidiomata of the saprobe/pathogen
Schizophyllum commune Fr. were collected, too [
7
]. It is a common cosmopolitan decomposer
of wood on rather dry, undecayed logs and stumps. This one was also collected in the
Czechish greenhouses (Paskow) [
11
]. In the Warsaw tropical greenhouses, on a dead
standing trunk of Washingtonia filifera (Linden ex Andre) H.Wendl, the cyphelloid saprobe
Henningsomyces candidus (Pers.) Kuntze, was found—a native species widely spread in
Europe as well as in North Africa, Asia, Australia, North America and South America [
10
].
Also, the corticoid saprobe Trechispora farinacea (Pers.) Liberta, was collected on a dead
part of the trunk of Cyathea australis Domin [
10
,
19
]. This fungal species is native and very
common in all the European countries, in Asia, Australia, North and South America [
15
].
In general, the lists of aphyllophoroid fungi in the greenhouses of individual cities include
only 2–6 species, which is 5–14 times less than the lists of agaricoid fungi. Such low results
are of interest due to the fact that aphyllophoroid fungi are among the most active wood
destroyers in natural ecosystems [13,15].
In Eastern Europe, there have been rare studies of macrofungi in greenhouses. In Saint
Petersburg, located in the boreal zone, in the greenhouses of the Peter the Great Botanical
Garden on non-native woody plants, on the soil under them, as well as on building
materials, structures, fences, shelves made of local woody plants, mainly local species of
agaricoid, aphyllophoroid and gasteroid macrofungi were found in 1920–1930 [
21
–
27
]. The
list of species is dominated by native fungal species, while non-native taxa are rare. At
the beginning of the 21st century, in the city of Ekaterinburg, in the greenhouses of the
J. Fungi 2023,9, 1116 3 of 23
Botanical Garden of the Ural Branch of the Russian Academy of Sciences, a number of
aphyllophoroid fungi were collected on non-native trees [
28
–
31
]. In general, the identified
species composition of macrofungi in the greenhouses of these two cities to 2010 was
extremely poor. For example, seven species of aphyllophoroid fungi on non-native plants
in Saint Petersburg and six species in Ekaterinburg were identified. However, even at
the beginning of the 20th century, it was assumed that despite the small lists of fungal
species in individual greenhouses, when summing up data from many greenhouses in Saint
Petersburg, the total number of species may not be inferior to the species richness in natural
conditions, when comparing similar areas of greenhouses and natural conditions [23].
According to the published results for the greenhouses of Central Europe, the identifi-
cation of fungal fruitbodies is random, many species are represented by single specimen,
and fungal diversity in greenhouses is lower compared to natural conditions [
7
,
10
,
19
].
However, the articles do not indicate what diversity level (alpha, beta or gamma) was
investigated. In this study, we test the hypothesis that fungal diversity in greenhouses is
poorer compared to natural conditions.
In order to find answers to the problems described above, over the last 20 years, our
team has been conducting research on the taxonomic diversity of aphyllophoroid fungi in
greenhouses in three cities of the boreal part of Eastern Europe: Saint Petersburg (the largest
complex of greenhouses is located in the Peter the Great Botanical Garden), Moscow (Tsitsin
Main Botanical Garden) and Ekaterinburg (several glasshouses including Botanical Garden
of UrB RAS). These greenhouses are of different ages and are also located in different
sectors of the continentality of the boreal climate: from the submaritime sector in Saint
Petersburg to the subcontinental one in Ekaterinburg.
The purpose of the study is to establish the number of aphyllophoroid fungi species
forming fruitbodies on non-native woody and herbaceous plants in greenhouses of Saint
Petersburg, Moscow and Ekaterinburg. The answer to this question will enable us to
estimate how poor aphyllophoroid fungal diversity in the boreal urban greenhouses com-
pared to natural condition. The following questions will also be explored: Do native or
non-native fungal species, saprobes or pathogens predominate? Are non-native or native
plant pathogens predominant? What species of fungi develop on structures made of local
wood? Are there non-native fungal species on them? Is the number of fungal species
related to the size, age of greenhouses, age or size of plants?
2. Materials and Methods
2.1. Study Area
The studied greenhouses are located in three cities of Eastern Europe: Saint Petersburg,
Moscow, and Ekaterinburg (Figure 1). The city of Saint Petersburg (60
◦
00
0
N, 30
◦
18
0
E;
12 m a.s.l.) has an area of 1439 km
2
with a population of 5.6 mL. The city is located at
the border of south boreal and hemiboreal vegetation subzones [
32
], with submaritime
climate, annual average precipitation 662 mm/year, air humidity 78%, and average annual
air temperature 6.6
◦
C [
33
]. Over the past 40 years, the average annual temperature in
the city has increased by 1.6
◦
C (in 1960–1990, it was 5.0
◦
C, and in 2000–2022, it reached
6
.6 ◦C
). The city of Moscow (55
◦
45
0
N, 37
◦
36
0
E; 165 m a.s.l.) has an area of 2561 km
2
and a
population of 13.1 mL. Located in the hemiboreal vegetation subzone, the annual average
precipitation is 708 mm/year, air humidity 78%, and the average annual air temperature
6.7
◦
C. Over 40 years, the average annual temperature has increased by 1.6
◦
C (from 5.1
◦
C
to 6.7
◦
C). The area of Ekaterinburg city (56
◦
48
0
N, 60
◦
35
0
E; 280 m a.s.l.) covers 568 km
2
,
whereas the population reaches 1.6 mL. The city is located at the border of south boreal
and hemiboreal vegetation subzones, on the border between Europe and Asia. The average
amount of precipitation is 562 mm per year, and the average air humidity is 71%. The
average annual air temperature is 3.6
◦
C, and has increased by 1.5
◦
C over 40 years (from
2.1
◦
C to 3.6
◦
C). The climate is continental with a rather characteristic sharp variability of
weather conditions and well-defined seasons [33].
J. Fungi 2023,9, 1116 4 of 23
J. Fungi 2023, 9, x FOR PEER REVIEW 4 of 23
nual air temperature is 3.6 °C, and has increased by 1.5 °C over 40 years (from 2.1 °C to 3.6
°C). The climate is continental with a rather characteristic sharp variability of weather con-
ditions and well-defined seasons [33].
Thus, the three studied cities are located in the boreal natural zone, but in different
sectors of the climate continentality: from the mild submaritime in Saint Petersburg,
through Moscow and to the subcontinental in Ekaterinburg with long and harsh winters.
Figure 1. Locations of the three studied cities in Eastern Europe: Saint Petersburg, Moscow, Ekate-
rinburg.
2.2. Description of Greenhouses in the Three Cities
In Saint Petersburg (SPB), the study was carried out in the largest complex of
greenhouses located in the territory of the Peter the Great Botanical Garden. The area of
these greenhouses is 1 ha (Table 1, Figure 2). In Moscow (MSC), the largest complex of
greenhouses located in the territory of the Main Botanical Garden, where the area of
greenhouses is 1 ha, was studied. In Ekaterinburg (EKB), a number of greenhouses lo-
cated in dierent parts of the city were studied. The largest area of greenhouses is located
in the territory of the Botanical Garden of the Ural Branch of the Russian Academy of
Sciences (0.3 ha), also in the territory of two city Arboreta (0.14 and 0.12 ha), and also in
the Botanical Garden of the Ural Federal University (0.05 ha). In total, the studied area of
greenhouses in Ekaterinburg is 0.6 ha.
The average age of greenhouses in the Peter the Great Botanical Garden (SPB) is 110
years, 40 years in the Main Botanical Garden (MSC) and 53 years in Ekaterinburg (EKB)
[34–36] (Table 1).
Table 1. Average parameters for greenhouses in Saint-Petersburg, Moscow and Ekaterinburg.
Greenhouses
Year of
Erection
Area, ha
Temperature in
Winter/Summer,
°C
Humidity in
Winter/Summer,
%
Major Collections
Peter the Great Botanical
Garden (SPB)
110
1.0
12–21/*
**/40–70
Musa, Punica, Coffea, Camellia,
Fuchsia, Bougainvillea, Dicksonia,
Cyathea, Cibotium, Fuchsia,
Plumeria, Begonia, Agathis
Main Botanical Garden
(MSC)
40
1.0
12–20/*
**/40–80
Nerium,Dicksonia, Cyathea,
Cibotium, Cycadopsida, Ficus,
Crinum, Hibiscus, Begonia, Cycas,
Passiflora, Anthurium, Pellea
●
●
●
Figure 1.
Locations of the three studied cities in Eastern Europe: Saint Petersburg, Moscow,
Ekaterinburg.
Thus, the three studied cities are located in the boreal natural zone, but in different
sectors of the climate continentality: from the mild submaritime in Saint Petersburg,
through Moscow and to the subcontinental in Ekaterinburg with long and harsh winters.
2.2. Description of Greenhouses in the Three Cities
In Saint Petersburg (SPB), the study was carried out in the largest complex of green-
houses located in the territory of the Peter the Great Botanical Garden. The area of these
greenhouses is 1 ha (Table 1, Figure 2). In Moscow (MSC), the largest complex of green-
houses located in the territory of the Tsitsin Main Botanical Garden, where the area of
greenhouses is 1 ha, was studied. In Ekaterinburg (EKB), a number of greenhouses located
in different parts of the city were studied. The largest area of greenhouses is located in the
territory of the Botanical Garden of the Ural Branch of the Russian Academy of Sciences
(0.3 ha), also in the territory of two city Arboreta (0.14 and 0.12 ha), and also in the Botanical
Garden of the Ural Federal University (0.05 ha). In total, the studied area of greenhouses in
Ekaterinburg is 0.6 ha.
Table 1. Average parameters for greenhouses in Saint-Petersburg, Moscow and Ekaterinburg.
Greenhouses Year of
Erection Area, ha
Temperature in
Winter/Summer,
◦C
Humidity in
Winter/Summer,
%
Major Collections
Peter the Great Botanical
Garden (SPB) 110 1.0 12–21/* **/40–70
Musa,Punica,Coffea,Camellia,
Fuchsia,Bougainvillea,Dicksonia,
Cyathea,Cibotium,Fuchsia,
Plumeria,Begonia,Agathis
Tsitsin Main Botanical
Garden (MSC) 40 1.0 12–20/* **/40–80
Nerium,Dicksonia,Cyathea,
Cibotium,Cycadopsida,Ficus,
Crinum,Hibiscus,Begonia,Cycas,
Passiflora,Anthurium,Pellea
Botanical Garden of the
UrB RAS (EKB) 53 0.61 13–16/* **/35–70
Citrus,Dicksonia,Cyathea,Musa,
Punica,Coffea,Camellia,Fuchsia,
Bougainvillea,Brunfelsia,Olea,
Platycerium,Camellia,Agave
Note: *—no heating, indoor temperature depends on external conditions, a system of blackout blinds is used in
hot weather; **—no fogging.
J. Fungi 2023,9, 1116 5 of 23
J. Fungi 2023, 9, x FOR PEER REVIEW 5 of 23
Botanical Garden of the
UrB RAS (EKB)
53
0.61
13–16/*
**/35–70
Citrus, Dicksonia, Cyathea, Musa,
Punica, Coffea, Camellia, Fuchsia,
Bougainvillea, Brunfelsia, Olea,
Platycerium, Camellia, Agave
Note: *—no heating, indoor temperature depends on external conditions, a system of blackout blinds
is used in hot weather; **—no fogging.
Figure 2. Greenhouses in the studied cities of Eastern Europe. (A,B)—Peter the Great Botanical
Garden (Saint-Petersburg); (C,D)—N.V. Tsitsin Main Botanical Garden (Moscow); (E,F)—Botanical
Garden of Ural Branch of the Russian Academy Sciences (Ekaterinburg).
Figure 2.
Greenhouses in the studied cities of Eastern Europe. (
A
,
B
)—Peter the Great Botanical
Garden (Saint-Petersburg); (
C
,
D
)—Tsitsin Main Botanical Garden (Moscow); (
E
,
F
)—Botanical Garden
of Ural Branch of the Russian Academy Sciences (Ekaterinburg).
The average age of greenhouses in the Peter the Great Botanical Garden (SPB) is 110
years, 40 years in the Tsitsin Main Botanical Garden (MSC) and 53 years in Ekaterinburg
(EKB) [34–36] (Table 1).
The composition of plants in the greenhouses of EKB has been studied in detail. All
considered plants are divided into two groups—woody and herbaceous. Woody plants
J. Fungi 2023,9, 1116 6 of 23
are subdivided into gymnosperms, angiosperms and arborescent. Herbaceous plants are
subdivided into evergreen perennials, ferns and annual grasses. The study takes into
account plants above 0.5 m in height. The age of each plant is also established. The results
of plant accounting were averaged over 10-year age classes: 0–9, 10–19, etc., until 110–119
years (oldest age class). Latin names of plants and names of taxon authors are taken from
the Plants of the World Online database [37].
2.3. Anthropogenic Gradient
Due to the fact that the study examines the issue of “poorness of the mycobiota in
greenhouses”, its testing was carried out in EKB. The anthropogenic gradient was studied
starting from the territory with the maximum proportion of non-native plants, i.e., from
greenhouses, in which 100% of the plants are non-native to the local flora. Further, the
proportion of non-native plants decreases in the Botanical Garden of the Ural Branch of
the Russian Academy of Sciences, plantations in the center of EKB, suburban forest parks,
and there are no non-native plants in old-growth forests in the vicinity of the city. As
regards old-growth forests, the Natural Park “Bazhov’s Tales” located 30 km from EKB
was used. The Botanical Garden, the center of Ekaterinburg and its environs are located
in forests dominated by Scotch pine (Pinus sylvestris L.), the same as within Ekaterinburg.
The duration of the study was 20 years (2003–2023). Fruitbodies of aphyllophoroid fungi
developing on woody and herbaceous local and non-native plants, saprobes, mycorrhizal
and phytopathogens were collected.
Thus, mycobiota of 5 plots were studied along the anthropogenic gradient:
1.
Greenhouses: A total of 8 greenhouses in EKB were investigated, the total area was
0.61 ha. The average area of greenhouses was 0.08 ha (from 0.05 to 0.12 ha). Thus, a
site of 0.08 ha was used as a model area. A total of 8 sites were studied in greenhouses.
Outside the greenhouses, in forests and parks, we also studied sites with an area of
0.08 ha, i.e., radius of 16 m.
2.
Botanical Garden of the Ural Branch of the Russian Academy of Sciences: 20 sites,
0.08 ha each.
3. City center of EKB: 20 sites, 0.08 ha each.
4. Suburban forest park: 20 sites, 0.08 ha each.
5. Old-growth forests in the vicinity of EKB: 20 sites, 0.08 ha each.
Therefore, a total of 88 sites were studied, 0.08 ha each.
2.4. Mycological Research
The fruitbodies of fungi found only on trees (live and dead, standing and fallen) and
roots were taken into account. Fungal species that formed fruitbodies on the soil were
excluded from the study. The term “specimen” means the detection of one fungal species
on a substrate (log, trunk, or branch) regardless of the number of fruitbodies that make up
such a specimen of the species found [
38
]. The specimen could consist of several fruitbodies.
Species richness of mycobiota per area unit was set as the average number of species per
hectare (species/ha). All analyzed fungal specimens were collected by the authors of this
article in the period of 2003–2023. We have added one species to the list (Flaviporus brownii
(Humb.) Donk) represented by one specimen which was collected in the greenhouses of the
Peter the Great Botanical Garden (SPB) in the 1930s by Prof. A.S. Bondartsev (depositing in
LE F), but we did not find this species in the SPB greenhouses.
In all greenhouses, fungi were collected using the route method [
38
] with the study
of the maximum possible number of plants. In order to study the hypothesis of how poor
the mycobiota diversity is in greenhouses, fungal specimens were collected within 88 sites
located in EKB greenhouses and model sites in Ekaterinburg city center and forested areas
in the suburbs (see Section 2.3). For specimen collection, the route method was used on
average 4 times during year over the past 20 years. The maximum range of tree and shrub
substrates located inside the sites was studied. The fruitbodies of fungi developing on
living trees were classified as pathogens, but if fruitbodies were collected on dead branches
J. Fungi 2023,9, 1116 7 of 23
and trunks, then they were classified as saprobes. Mycorrhiza-formers included species
classified according to UNITE (https://unite.ut.ee/ (accessed on 23 June 2023)).
All fungal specimens collected by the authors were identified based on the morpho-
logical features of fruitbodies and according to the following key books [
13
,
15
,
27
,
39
], using
light microscopes LEICA 2000 (Wetzlar, Germany) and ZEISS Axio Imager A1 (Jena, Ger-
many). In our studies, we adhered to a classical morphological approach, based on the
idea that the development of fruitbodies marks the accumulation of mycelium biomass
within the substrate, i.e., predominance of this species in the corresponding substrate entity.
We agree that molecular screening can reveal greater species diversity per substrate entity,
but it often also reveals inactive propagules and ephemeral mycelia of low competitive
potential, i.e., incapable of its complete colonization [
40
]. The collected specimens are
deposited in the Institute of Plant and Animal Ecology Herbarium (SVER) and Komarov
Botanical Institute Herbarium (LE F). The fungal species nomenclature is given according
to Index Fungorum [41].
All fungal species were subsumed under one of the two categories: native or non-
native. Non-native species were defined as species untypical of the local mycobiota, whose
introduction into a given area was not associated with the natural course of mycogenesis,
but resulted from direct or indirect human activity [
2
]. The main source used to distinguish
between native and non-native species was Aphyllophoroid fungi of Sverdlovsk region [
31
].
The GBIF data on the worldwide distribution of fungal species (www.gbif.org; accessed on
19 April 2023) were used.
2.5. Statistical Analysis
In order to draw a dendrogram reflecting the similarity of the species composition
of fungi along the anthropogenic gradient, the Ward method and the Pearson correlation
coefficient were used. The Spearman’s rank correlation coefficient (r) was used to establish
a linear correlation between various parameters of mycobiota, climate, and plants.
The
α
-diversity of mycobiota was estimated in three ways: as the average number
of fungal species in five plots along anthropogenic gradient as well as by the value of the
Shannon index, and the Menhinick index. The
β
-diversity was also assessed in three ways:
the average Jaccard index as well as the Morisita–Horn Index and the Whittaker Index [
38
].
Statistical differences between the sites along the anthropogenic gradient were established
using the Mann–Whitney (U-test) method.
An approach based on the sampling algorithm was used to estimate the expected
number of species per transect [
42
]. This approach is based on a rarefaction curve drawn
using a special algorithm for random multiple permutation of data within samples from
among the detected samples. An indirect method for assessing the completeness of the
identification of species richness was also used—the Turing coefficient (C), which was
based on the ratio between the number of singleton species (represented by a single find)
and the total number of identified species [43].
C = (1 −f1/S) ×100%, (1)
where f
1
is the number of singleton species, and S is the total number of identified species.
The potential number of species (T) can be calculated as
T = S/C (2)
Statistical data processing was carried out using statistical software packages Statistica
10.0 and MS Excel 2007.
3. Results
3.1. List of Fungal Species in the Greenhouses of the Three Cities
During this study, 140 samples representing 58 species of aphyllophoroid macrofungi
(Table S1) were collected in the greenhouses of the three cities (Table 2). Most specimens
J. Fungi 2023,9, 1116 8 of 23
and species were collected in the greenhouses of EKB (84 specimens/39 species), then in
the Main Botanical Garden of MSC (39/28), and to a lesser extent in the Peter the Great
Botanical Garden of SPB (17/17).
Table 2.
Species list of aphyllophoroid fungi in the greenhouses of the three boreal cities (number
of specimens).
Species Trophic
Group
City
Information
SPB MSC EKB
Abortiporus biennis
(Bull.) Singer Sap 1 EKB: at the base of Dypsis lutescens (H. Wendl.) Beentje et
J.Dransf. (Arecaceae) and on woody box.
Antrodia gossypium
(Speg.) Ryvarden Sap 1 1 3
SPB: on dead part of deciduous tree/MSC: at the base of Ginkgo
biloba L. (Ginkgoaceae)/EKB: at the base of Cryptomeria japonica
(Thunb. ex L.f.) D.Don (Cupressaceae); on Ficus carica L.
(Moraceae); on dead part of Eriobotrya japonica (Thunb.)
Lindl. (Rosaceae)
Athelia bombacina
(Link) Pers. Sap 1 MSC: on dead part of Cycas revoluta Thunb. (Cycadaceae)
Athelia epiphylla Pers. Sap 2 EKB: on dead petioles of Pteris sp. (Pteridiaceae); on root neck
of Strelitzia reginae Banks ex Aiton (Strelitziaceae).
Athelia rolfsii (Curzi)
C.C. Tu et Kimbr. Pat 1 EKB: at the base of Hedera helix L. (Araliaceae).
Bjerkandera adusta
(Willd.) P. Karst. Sap 2 2
MSC: at the base of Berberis rotundifolia Poepp. et Endl.
(Berberiaceae); on dead stem of Eleodendron croceum (Thunb.)
DC. (Celastraceae)/EKB: dead branch of Laurus nobilis L.
(Lauraceae); at the base of Ceratonia siliqua L. (Fabaceae).
Botryobasidium
conspersum J. Erikss. Sap 1 SPB: on dead branch of Vitis vinifera L. (Vitaceae).
Brevicellicium olivascens
(Bres.) K.H. Larss. et
Hjortstam
Sap 1 EKB: at the base of Pteris sp. (Pteridiaceae).
Byssomerulius corium
(Pers.) Parmasto Sap 1 1
SPB: at the base of Hedera helix L. (Araliaceae)/MSC: at the base
of Quercus acuta Thunb. (Fagaceae).
Ceratobasidium
cornigerum (Bourdot)
D.P. Rogers
Sap 1 2
MSC: at the base of Cordyline banksii Hook f.
(Asparagaceae)/EKB: base of Musa basjoo Siebold et Zucc. ex
Iinuma (Musaceae); base of Prunus laurocerasus L. ( Rosaceae ).
Cerioporus varius (Pers.)
Zmitr. et Kovalenko Sap 1 SPB: on dead part of deciduous tree.
Ceriporia viridans (Berk.
et Broome) Donk Sap 1
EKB: on dead branch of Citrus
×
sinensis (L.) Osbeck (Rutaceae).
Clavulina cinerea (Bull.)
J. Schröt. Myc 1 SPB: at the base and on the roots of alive Nerium oleander L.
(Apocynaceae).
Clavulina ornatipes
(Peck) Corner Myc 1 EKB: on the roots of Magnolia obovata Thynb. (Magnoliaceae).
Clavulinopsis
aurantiocinnabarina
(Schwein.) Corner
Sap 1 EKB: on the roots of Magnolia obovata Thunb. (Magnoliaceae)
and Cryptomeria japonica (Thunb. ex L.f.) D.Don (Cupressaceae).
Coniophora arida (Fr.) P.
Karst. Sap 1 2
MSC: at the base of Ginkgo biloba L. (Ginkgoaceae)/EKB: dry
base of Cereus repandus Mill. (Cactaceae); at the base of
Ligustrum ovalifolium Hassk. (Oleaceae).
J. Fungi 2023,9, 1116 9 of 23
Table 2. Cont.
Species Trophic
Group
City
Information
SPB MSC EKB
Coniophora puteana
(Schumach.) P. Karst. Sap 2 3
MSC: at the base of Pittosporum tobira (Thunb.) W.T.Aiton
(Pittosporaceae); dead part of Cordyline pumilio Hook.f.
(Asparagaceae)/EKB: at the base of Cupressus sempervirens L.
(Cupressaceae); dead base of Pteris sp. (Pteridiaceae); base of
Dendrobium nobile Lindley (Orchidaceae).
Cylindrobasidium
evolvens (Fr.) Jülich Sap 2 5
MSC: broken branch of Acca sellowiana (O.Berg) Burret
(Myrtaceae); at the base of fern (Pteridiaceae)/EKB: on dead
stem of Hedera helix L. (Araliaceae); dead leave of Agave
americana L. (Asparagales); dead branch of Bougainvillea
spectabilis Willd. (Nyctaginaceae); dead branch of Hydrangea
macrophylla (Thunb.) Ser. (Hydrangeaceae); dead petioles of
Cycas revoluta Thunb. (Cycadaceae).
Flaviporus brownii
(Humb.) Donk Sap 1 SPB: on dead deciduous tree and box wall (Bondartsev, 1953).
Ganoderma applanatum
(Pers.) Pat. Sap 1 EKB: at the base of Coffea arabica L. (Rubiaceae).
Helicobasidium
purpureum (Tul.) Pat. Pat 1
MSC: at the base of Asparagus falcatus (L.) Druce (Asparagales).
Hymenochaete rheicolor
(Mont.) Lév. Sap 1 MSC: at the base of Sparmannia africana L.f. (Malvaceae).
Hymenochaete tabacina
(Fr.) Lev. Sap 1 1
SPB: at the base of deciduous tree/MSC: on dead part of Citrus
reticulata Blanco (Rutaceae).
Hyphodontia arguta (Fr.)
J. Erikss. Sap 1 2 3
SPB: on dead branch of Citrus ×sinensis (L.) Osbeck
(Rutaceae)/MSC: on the leaves of Sabal palmetto (Walter) Lodd.
ex Schult. et Schult.f. (Arecaceae) ; at the base of Ficus lyrata
Warb. (Moraceae)/EKB: dead part of Monstera deliciosa Liebm.
(Araceae); on rotten part of Lepidozamia peroffskyana Regel
(Zamiaceae); on dead stem of Hedera helix L. (Araliaceae).
Hypochniciellum
ovoideum (Jülich)
Hjortstam
Sap 1 EKB: at the base of Tetrastigma voinierianum
(Sallier) Pierre ex Gagnep. (Vitaceae).
Lentinus brumalis
(Pers.) Zmitr. Sap 1 1 SPB: dead branch of Laurus nobilis L. (Lauraceae)/MSC: at the
base of Acca sellowiana (O.Berg) Burret (Myrtaceae).
Leptosporomyces
raunkiaeri (M.P. Christ.)
Jülich
Sap 1 EKB: at the base and dead roots of Chamaerops humilis L.
(Arecaceae).
Lindtneria trachyspora
(Bourdot et Galzin)
Pilát
Sap 1 MSC: at the base of Drimys granadensis L.f. (Winteraceae).
Lyomyces crustosus
(Pers.) P. Karst. Sap 4
EKB: on dead stem of Hedera helix L. (Araliaceae); at the base of
Pteris sp. (Pteridiaceae); dead stem of Monstera deliciosa Liebm.
(Araceae); dead base of Aspedistra lurida Ker Gawl.
(Asparagaceae).
Lyomyces erastii
(Saaren. et Kotir.)
Hjortstam et Ryvarden
Sap 2 3
MSC: on dead leaf of Washingtonia robusta H. Wendl.
(Arecaceae); on dead stem of Vitis vinifera L. (Vitaceae)/EKB:
dead branch of at the base of Ginkgo biloba L. (Ginkgoaceae);
dead root of Philodendron bipinnatifidum Schott ex Endl.
(Araceae); on dead branch of Psidium guajava L. (Myrtaceae).
J. Fungi 2023,9, 1116 10 of 23
Table 2. Cont.
Species Trophic
Group
City
Information
SPB MSC EKB
Lyomyces sambuci
(Pers.) P. Karst. Sap 1 3 6
SPB: at the base of deciduous tree/MSC: on dead branch of
Elaeodendron capense Eckl. et Zeyh. (Celastraceae); dead branch
of Citrus maxima (Burm.) Merr. ( Rutaceae); dead base of Clivia
miniata (Lindl.) Regel. (Amaryllidaceae)/EKB: On dead twig of
Citrus medica L. (Rutaceae); on dead branch of Acca sellowiana
(O.Berg) Burret (Myrtaceae); at the base of Ficus carica L.
(Moraceae); at the base of Musa maclayi F.Muell. ex
Mikl.-Maclay (Musaceae); on dead petioles of Blechnum
brasiliense Desv. (Blechnaceae); on dead base of Strelitzia reginae
Banks ex Aiton (Strelitziaceae).
Microporus xanthopus
(Fr.) Kuntze Sap 1
EKB: on woody box made of East-Asian Carpinus cordata Blume
(Betulaceae) where grew Magnolia obovata Thunb.
(Magnoliaceae).
Peniophora cinerea
(Pers.) Cooke Sap 1 3 5
SPB: on twig of deciduous tree/MSC: on branch of Laurus
novocanariensis Rivas Mart., Lousa, Fern. Prieto, E.Diaz,
J.C.Costa et C.Aguiar (Lauraceae); on dead branch of Eucalyptus
oblique L’Her. (Myrtales); on dead part of Paphiopedilum
haynaldianum (Rchb.f.) Stein (Orchidaceae)/ EKB: on dead stem
of Vitis vinifera L. (Vitaceae); on dead twig of Citrus limon (L.)
Osbeck. (Rutaceae); on dead branch of Ficus carica L.
(Moraceae); on dead stem of Aloe arborescens Mill.
(Asphodelaceae); on dead stem of Heptapleurum arboricola
Hayata (Araliaceae).
Physalacria cryptomeriae
Berthier et Rogerson Sap 1 EKB: on dead twigs of Cryptomeria japonica (Thunb.ex L.f.)
D.Don (Cupressaceae).
Physalacria orientalis
(Kobayasi) Berthier Sap 2
EKB: on dead branches and rotten base of Magnolia grandiflora L.
(Magnoliaceae); at the base of Grevillea robusta A.Cunn. ex R.Br.
(Proteaceae).
Piloderma byssinum (P.
Karst.) Jülich Myc 2 3
MSC: at the base of Ocotea foetens (Aiton) Baill. (Lauraceae);
dead branch of Nerium oleander L. (Apocynaceae)/EKB: at the
base of Cryptomeria japonica (Thunb.ex L.f.) D.Don
(Cupressaceae); at the base of Phoenix roebelenii T.Anderson
(Arecaceae); on the rotten base of Iris japonica Thunb.
(Iridaceae).
Ramariopsis minutula
(Bourdot et Galzin)
R.H. Petersen
Sap 1 MSC: on the roots of Dicksonia fibrosa Colenso (Dicksoniaceae).
Ramariopsis kunzei (Fr.)
Corner Sap 1 1 2
SPB: on the roots of Blechnum brasiliense Desv.
(Blechnaceae)/MSC: on the roots and at the base of Olearia
tomentosa (J.C.Wendl.) DC. (Ateraceae)/EKB: on the roots and
at the base of Abutilon darwinii Hook.f. (Malvaceae); on the
roots of Ilex crenata Thunb. (Aquifoliaceae).
Rhizoctonia fusispora (J.
Schröt.) Oberw., R.
Bauer, Garnica et R.
Kirschner
Sap 1 2
MSC: on the roots and bark of Petrea volubilis L.
(Verbenaceae)/EKB: at the base of Taxus baccata L. (Taxaceae);
on the soil and roots of Casuarina equisetifolia L. (Casuarinaceae).
Rigidoporus lineatus
(Pers.) Ryvarden Sap 1
EKB: at the base of Eucalyptus sp. (Myrtaceae) and the wall of a
clay pot brought in 1958 from Batumi Botanical Garden.
Fruitbodies were collected in 2002.
Scopuloides rimosa
(Cooke) Jülich Sap 1 SPB: at the base of deciduous tree.
J. Fungi 2023,9, 1116 11 of 23
Table 2. Cont.
Species Trophic
Group
City
Information
SPB MSC EKB
Sistotrema brinkmannii
(Bres.) J. Erikss. Sap 2
EKB: at the base of Casuarina equisetifolia L. (Casuarinaceae); at
the base of Musa basjoo Siebold and Zucc. ex Iinuma
(Musaceae).
Sertulicium
niveocremeum (Höhn.
et Litsch.) Spirin et
K.H. Larss.
Sap 2
EKB: at the base of Grevillea robusta A.Cunn. ex R.Br.
(Proteaceae); at the base of Cinnamomum verum J.Presl.
(Lauraceae).
Subulicystidium
longisporum (Pat.)
Parmasto
Sap 1 SPB: on rotten part of Vitis vinifera L. (Vitaceae).
Thanatephorus
terrigenus (Bres.)
G.Langer
Sap 1 MSC: at the base of Meryta sinclairii (Hook.f.) Seem.
(Araliaceae).
Thelephora palmata
(Scop.) Fr.
Myc/Pat 1
MSC: at the base and roots of Myrsine africana L. (Primulaceae).
Thelephora terrestris
Ehrh. ex Fr.
Myc/Pat 1 SPB: at the base of deciduous tree.
Tomentella cinerascens
(P. Karst.) Höhn. et
Litsch.
Myc 1
MSC: at the base of Acca sellowiana (O.Berg) Burret (Myrtaceae).
Tomentella lapida (Pers.)
Stalpers Myc 1 EKB: at the base of Magnolia grandiflora L. (Magnoliaceae).
Tomentella radiosa (P.
Karst.) Rick Myc 2
EKB: at the base of Manihot esculenta Crantz. (Euphorbiaceae);
at the base of Musa basjoo Siebold et Zucc. ex Iinuma
(Musaceae).
Trametes ochracea
(Pers.) Gilb. et
Ryvarden
Sap 1 2
MSC: at the base of Uncarina grandidieri (Baill.) Stapf
(Pedaliaceae)/EKB: at the base of Ginkgo biloba L. (Ginkgoaceae);
at the base of Ficus carica L. (Moraceae).
Trechispora nivea (Pers.)
K. H. Larss. Sap 1 SPB: at the base of fern.
Trechispora farinacea
(Pers.) Liberta Sap 1 1 2
SPB: at the base of Dicksonia fibrosa Colenso
(Dicksoniaceae)/MSC: at the base of Eriobotrya japonica (Thunb.)
Lindl. (Rosaceae)/EKB: at the base of Myriocarpa cordifolia
Liebm. (Urticaceae); on the dead base of Zingiber officinale
Roscoe (Zingiberaceae).
Typhula micans (Pers.)
Berthier Sap 1 2
MSC: at the dead base of Dianella sp. (Asphodelaceae)/EKB: on
dead part of Brasiliopuntia brasiliensis (Wild.) A.Berger
(Cactaceae); on dead stem of Sonchus canariensis (Sch. Bip.)
Boulo (Asteraceae).
Xylodon asper (Fr.)
Hjortstam et Ryvarden
Sap 2
EKB: at the base of Agave tequilana F.A.Webber (Asparagaceae);
at the base of Tetrastigma voinierianum (Sallier) Pierre ex Gagnep.
(Vitaceae).
Xylodon detriticus
(Bourdot) K.H. Larss.,
Viner et Spirin
Sap 3
EKB: on dead base of Strelitzia nicolai Regel et K.Koch
(Strelitziaceae); at the base of Ficus carica L. (Moraceae); on dead
base of Ruscus aculeatus L. (Asparagaceae).
J. Fungi 2023,9, 1116 12 of 23
Table 2. Cont.
Species Trophic
Group
City
Information
SPB MSC EKB
Xylodon flaviporus
(Berk. et M.A. Curtis
ex Cooke) Riebesehl et
Langer
Sap 2 5
MSC: on dead parts of Distilium racemosum Siebold et Zucc.
(Hamamelidaceae); at the base of Barberis rotundifolia
(Rosaceae)/EKB: on dead branch of Carica papaya L.
(Caricaceae); on dead part of Cereus repandus Mill. (Cactaceae);
on dead branch of Nolina recurvata Lem. (Asparagaceae); on
dead stem of Ficus elastic Roxb. ex Hornem. (Moraceae); at the
base of Cycas revoluta Thunb. (Cycadaceae).
Xylodon raduloides
Riebesehl et Langer Sap 2 EKB: on dead roots of Monstera deliciosa Liebm. (Araceae); on
dead stem of Pancratium L. (Amaryllidaceae).
Total: 17 39 84
Note: SPB—Saint Petersburg; MSC—Moscow; EKB—Ekaterinburg; Sap—saprobes; Myc—mycorrhizal;
Pat—pathogens.
An amount of 46 species (79.3% of the total) are local, widespread in the boreal part
of Eastern Europe, while 12 species (20.7%) have range centers in other biogeographic
regions of the planet. A number of non-native species at the time of their collection
were new to Russia. For example, in the 1920s, it was Flaviporus browni, and, in the 2000,
Physalacria cryptomeriae. In this paper, Hymenochaete rheicolor is reported in Russia for the
first time. In the greenhouses of SPB, one species can be classified as non-native (5.9% of the
number of species in SPB), in MSC, three species (10.7%), and, in EKB, eight (19.5%). Thus,
as the number of identified fungal species grows, the proportion of non-native species
increases linearly.
Poroid fungus Flaviporus brownii is non-native to Eastern Europe, collected in the palm
greenhouse of SPB [
27
] and widely spread in nemoral and subtropical Europe, sub/tropics
of America, Asia, Africa, Australia and New Zealand (https://www.gbif.org/ru/species/
5954717 (accessed on 19 June 2023)).
In MSC, a non-native poroid Lindtneria trachyspora was collected having a mode of
finds in nemoral and subtropical Europe and eastern North America; there were finds
in the tropics of Central and South America (Brazil, Costa Rica, El Salvador, Mexico),
Africa (Seychelles, South Africa), in Australia and East Asia (Japan) (https://www.gbif.
org/ru/species/2552491 (accessed on 19 June 2023). Stereoid fungus Hymenochaete rheicolor
has a mode of findings in the sub/tropical regions of America (Argentina, Brazil, Chile,
Costa Rica, Cuba, Dominican Republic, Ecuador, French Guiana, Mexico, Panama, Puerto
Rico, Jamaica, and the USA), Asia (China, India, Nepal, Philippines, Japan, and Thailand)
and Australia (https://www.gbif.org/en/species/2519646 (accessed on 19 June 2023).
Thanatephorus terrigenus is a European species known in Germany, Great Britain, Norway,
Sweden (https://www.gbif.org/ru/species/2555158 (accessed on 19 June 2023) and the
Urals [31].
In the EKB greenhouses, the poroid Microporus xanthopus was collected on the wall of
a box made of the wood of the East Asian Carpinus cordata Blume, in which the Magnolia
obovata Thunb. seedling had been brought from the Russian Far East [
44
]. Under the
natural conditions of Russia, this fungus was found only in the south of the Far East in
Primorsk Territory [
45
]. East Asian clavarioid fungi Physalacria cryptomeriae and tropical Ph.
orientalis were collected on dead branches of Cryptomeria japonica (Thunb. ex L.f.) D.Don.
and Magnolia grandiflora L., respectively. At the base of Eucalyptus sp. and on the wall
of a clay tub, a poroid Rigidoporus lineatus was collected—widespread in tropical regions,
but also often found in temperate greenhouses. An interesting finding was the corticoid
Leptosporomyces raunkiaeri known in Leningrad [
46
] and Nizhny Novgorod provinces of
Russia (https://www.gbif.org/ru/species/2554208 (accessed on 19 June 2023). This fungus
was recently found in Sverdlovsk province on a dead tree of Alnus glutinosa (L.) Gaertn.
in a suburban forest of EKB [
47
]. The question arises: did this fungus inhabit the natural
J. Fungi 2023,9, 1116 13 of 23
biocenoses of the region and from there “come“ to the greenhouses, or vice versa—was it
brought into the greenhouses, and is it now settling into nature?
In the three cities studied, there is no correlation between plant height and the number
of identified fungal species (r = 0.44, p= 0.37). It was also found that the number of fungal
species did not depend on the area of greenhouses (r = –0.85, p= 0.31) nor on the average
age of greenhouses (r = –0.86, p= 0.09). At the same time, in the EKB greenhouses, a strong
positive linear regression (Figure 3) was found between the level of sampling effort (as the
distance traveled when detecting fungi in greenhouses) with the number of detected fungal
specimens (r = 0.99, p= 0.0001) as well as polynomial fit (3 order) between the sampling
effort with the number of fungal species (R2= 0.96, p= 0.0002).
J. Fungi 2023, 9, x FOR PEER REVIEW 13 of 23
Britain, Norway, Sweden (https://www.gbif.org/ru/species/2555158 (accessed on 19 June
2023) and the Urals [31].
In the EKB greenhouses, the poroid Microporus xanthopus was collected on the wall
of a box made of the wood of the East Asian Carpinus cordata Blume, in which the Mag-
nolia obovata Thunb. seedling had been brought from the Russian Far East [44]. Under the
natural conditions of Russia, this fungus was found only in the south of the Far East in
Primorsk Territory [45]. East Asian clavarioid fungi Physalacria cryptomeriae and tropical
Ph. orientalis were collected on dead branches of Cryptomeria japonica (Thunb. ex L.f.)
D.Don. and Magnolia grandiora L., respectively. At the base of Eucalyptus sp. and on the
wall of a clay tub, a poroid Rigidoporus lineatus was collected—widespread in tropical re-
gions, but also often found in temperate greenhouses. An interesting nding was the
corticoid Leptosporomyces raunkiaeri known in Leningrad [46] and Nizhny Novgorod
provinces of Russia (hps://www.gbif.org/ru/species/2554208 (accessed on 19 June 2023).
This fungus was recently found in Sverdlovsk province on a dead tree of Alnus glutinosa
(L.) Gaertn. in a suburban forest of EKB [47]. The question arises: did this fungus inhabit
the natural biocenoses of the region and from there “come“ to the greenhouses, or vice
versa—was it brought into the greenhouses, and is it now seling into nature?
In the three cities studied, there is no correlation between plant height and the
number of identied fungal species (r = 0.44, p = 0.37). It was also found that the number
of fungal species did not depend on the area of greenhouses (r = –0.85, p = 0.31) nor on the
average age of greenhouses (r = –0.86, p = 0.09). At the same time, in the EKB greenhouses,
a strong positive linear regression (Figure 3) was found between the level of sampling
eort (as the distance traveled when detecting fungi in greenhouses) with the number of
detected fungal specimens (r = 0.99, p = 0.0001) as well as polynomial t (3 order) between
the sampling eort with the number of fungal species (R2 = 0.96, p = 0.0002).
Figure 3. Relationship between sampling effort (distance traveled within the greenhouses) and (A)
number of fungal specimens (y = 9.89x + 0.38, r = 0.99, p = 0.0001) and with (B) number of fungal spe-
cies (y = 0.05x3 − 1.24x2 +11.54x + 0.11, R2 = 0.99, p = 0.0002).
With the current number, age and condition of plants in EKB greenhouses, 52.1 ± 4.8
species of aphyllophoroid fungi can be identied (Figure 4) with 39 species collected by
us (75% of the potential species composition). Consequently, the revealed diversity is
currently far from the potentially possible one. At the same time, 12 out of 39 species of
aphyllophoroid fungi in the EKB greenhouses are characterized by a single nd (single-
tons); therefore, the Turing coecient is 69%, which indirectly conrms the estimated
species richness of 56 species.
0
20
40
60
80
100
0246810
number of specimens
distance, km
0
5
10
15
20
25
30
35
40
45
0 2 4 6 8 10
number of species
distance, km
A
B
Figure 3.
Relationship between sampling effort (distance traveled within the greenhouses)
a
nd (A) numb
er of fungal specimens (y = 9.89x + 0.38, r = 0.99, p= 0.0001) and with (
B
) number of
fungal species (y = 0.05x3−1.24x2+11.54x + 0.11, R2= 0.99, p= 0.0002).
With the current number, age and condition of plants in EKB greenhouses,
52.1 ±4.8
species of aphyllophoroid fungi can be identified (Figure 4) with 39 species collected by
us (75% of the potential species composition). Consequently, the revealed diversity is
currently far from the potentially possible one. At the same time, 12 out of 39 species of
aphyllophoroid fungi in the EKB greenhouses are characterized by a single find (singletons);
therefore, the Turing coefficient is 69%, which indirectly confirms the estimated species
richness of 56 species.
Six species of aphyllophoroid fungi (10.5% of the total number) were collected in
the greenhouses of all the three cities: Antrodia gossypium,Hyphodontia arguta,Lyomyces
sambuci,Peniophora cinerea,Ramariopsis kunzei, and Trechispora farinacea. All these species
were saprobes. Between the SPB and MSK greenhouses, the Jaccard similarity index is 0.27,
between SPB and EKB, it is 0.11, and, between MSK and EKB, it is 0.52.
Four species (6.9%) are phytopathogens (facultative). Three of them (Helicobasidium
purpureum,Thelephora palmata, and Th. terrestris) are local species widespread in nature
and greenhouses. One species, Athelia rolfsii, is non-native, with a mode of finds in the
sub/tropical regions of the planet, but has long been widely distributed in greenhouses
of the temperate zone (https://www.gbif.org/ru/species/2554082 (accessed on 23 June
2023)). Each of the four species of phytopathogens was collected in the greenhouses of only
one city.
J. Fungi 2023,9, 1116 14 of 23
J. Fungi 2023, 9, x FOR PEER REVIEW 14 of 23
Figure 4. Completeness of identification of aphyllophoroid fungi species in the EKB greenhouses for
20 years, depending on the number of specimens. A smoothed rarefaction curve (individual-based
rarefaction curve) is presented, depending on the number of specimens.
Six species of aphyllophoroid fungi (10.5% of the total number) were collected in the
greenhouses of all the three cities: Antrodia gossypium, Hyphodontia arguta, Lyomyces sam-
buci, Peniophora cinerea, Ramariopsis kunzei, and Trechispora farinacea. All these species were
saprobes. Between the SPB and MSK greenhouses, the Jaccard similarity index is 0.27,
between SPB and EKB, it is 0.11, and, between MSK and EKB, it is 0.52.
Four species (6.9%) are phytopathogens (facultative). Three of them (Helicobasidium
purpureum, Thelephora palmata, and Th. terrestris) are local species widespread in nature and
greenhouses. One species, Athelia rolfsii, is non-native, with a mode of finds in the
sub/tropical regions of the planet, but has long been widely distributed in greenhouses of
the temperate zone (https://www.gbif.org/ru/species/2554082 (accessed on 23 June 2023)).
Each of the four species of phytopathogens was collected in the greenhouses of only one
city.
There were 115 specimens collected on woody plants (82.1% of total), which com-
prised 50 fungal species (Table 3). These were mainly saprobes (42 species/102 specimens)
and mycorrhizal (4/9). All phytopathogenic fungi were collected on woody plants only. An
amount of 25 specimens of 10 species were collected on herbaceous plants. All species on
herbaceous plants were saprobes. On all plants, the largest number of specimens (90.7%)
belonged to saprobes, 6.4% to mycorrhizal, pathogens were represented by 2.8% of the
specimens. Fungi were not detected on herbaceous annual plants. Consequently, the
number of fungal specimens collected on woody plants exceeded the number on herba-
ceous ones in SPB by 16 times, in MSC by 5.3 times, in EKB by 3.4 times, and the average
for the three cities was 4.3 (Table 2). An amount of 88.9% of mycorrhizal fungi were col-
lected on angiosperms, while 52% of saprobes were collected on angiosperms, and 21.3%
on arborescent.
In the three surveyed cities, during 20 years, no fungal species was found twice or
more times on the same plant species. Moreover, not a single fungal species has been found
twice or more times on the same genus or even family of plants.
0
10
20
30
40
020 40 60 80 100
number of species
number of specimens
Figure 4.
Completeness of identification of aphyllophoroid fungi species in the EKB greenhouses for
20 years, depending on the number of specimens. A smoothed rarefaction curve (individual-based
rarefaction curve) is presented, depending on the number of specimens.
There were 115 specimens collected on woody plants (82.1% of total), which comprised
50 fungal species (Table 3). These were mainly saprobes (42 species/102 specimens) and
mycorrhizal (4/9). All phytopathogenic fungi were collected on woody plants only. An
amount of 25 specimens of 10 species were collected on herbaceous plants. All species
on herbaceous plants were saprobes. On all plants, the largest number of specimens
(90.7%) belonged to saprobes, 6.4% to mycorrhizal, pathogens were represented by 2.8%
of the specimens. Fungi were not detected on herbaceous annual plants. Consequently,
the number of fungal specimens collected on woody plants exceeded the number on
herbaceous ones in SPB by 16 times, in MSC by 5.3 times, in EKB by 3.4 times, and the
average for the three cities was 4.3 (Table 2). An amount of 88.9% of mycorrhizal fungi
were collected on angiosperms, while 52% of saprobes were collected on angiosperms, and
21.3% on arborescent.
Table 3.
Aphyllophoroid fungi number of specimens collected on woody and herbaceous plants in
the three urban greenhouses (number of specimens/%).
Parameter Fungal Trophic Group Total
Saprobes Pathogens Mycorrhizal
Plants
Woody 102/72.8 4/2.8 9/6.4 115/82.1
Gymnosperms
9/6.4 0/0 0/0 9/6.4
Angiosperms 66/14.1 2/1.4 8/5.7 75/53.6
Arborescent 27/19.3 2/1.4 1/0.7 30/21.4
Herbaceous 25/17.8 0/0 0/0 25/17.8
Evergreen 15/10.7 0/0 0/0 15/10.7
Ferns 10/7.1 0/0 0/0 10/7.1
Annual 0/0 0/0 0/0 0/0
Total 127/90.7 4/2.8 9/6.4 140/100
Note: If a fungus belongs to several trophic groups, then the species is taken into account in all corresponding
trophic groups in the table. The largest parameters are highlighted in bold.
J. Fungi 2023,9, 1116 15 of 23
In the three surveyed cities, during 20 years, no fungal species was found twice or
more times on the same plant species. Moreover, not a single fungal species has been found
twice or more times on the same genus or even family of plants.
3.2. Fungal Diversity on Model Plots in Ekaterinburg along the Anthropogenic Gradient
Taking the example of the EKB greenhouses as the best studied ones of the three
analyzed cities (the maximum sampling effort was applied, the most specimens and species
of fungi were collected), it is possible to study fungal diversity along the anthropogenic
gradient: from greenhouses, with a 100% set of non-native plants, through the Botanical
Garden UrB RAS, the center of EKB, suburban forest parks and old-growth forests located
40 km from Ekaterinburg, where only native plants grow (Table 4).
Table 4. Parameters of five plots along anthropogenic gradient in Ekaterinburg and surroundings.
Parameter Plot
Greenhouses
Arboretum of
Botanical Garden
UrB RAS
City Center Suburbs Forests Old-Growth
Forests
Number of sites 8 20 20 20 20
Average size of site, ha
0.08 0.08 0.08 0.08 0.08
Total size, ha 0.61 1.55 1.62 1.51 1.65
Share of non-native
tree species, % 100 88 (100–72) 73 (100–48) 42 (79–26) 0
Total number of
fungal species 39 86 43 64 68
Average number of
fungal species per site 5.9 (2–8) 18.3 (13–25) 7.7 (4–12) 15.8 (12–21) 18.1 (11–24)
Menhinick Index 4.19 3.53 4.02 4.99 5.58
Shannon Index 3.48 3.31 3.54 3.88 4.42
Jaccuard Index 0.08 (0–0.24) 0.38 (0.25–0.46) 0.32 (0.24–0.40) 0.57 (0.49–0.68) 0.62 (0.58–0.83)
Whittaker Index 5.6 3.7 4.6 3.0 2.7
Morisita-Horn Index 0.04 0.19 0.18 0.24 0.30
Total number of
non-native
fungal species
8 9310
Share of non-native
fungal species, % 20.5 9.5 6.9 1.6 0
Species richness of
non-native fungi, ha 13.1 5.8 1.8 0.7 0
Shown in parentheses is min.–max.
Three studied parameters of mycobiota
α
-diversity are lower in greenhouses compared
to natural conditions. For example, the average number of fungal species within a site is
3.0 ti
mes lower compared to natural conditions, the Menhinick index is 1.3 times lower, and
the Shannon index is 1.2 times lower. On the other hand, all three parameters reflecting the
β
-diversity of mycobiota are higher in greenhouses: the Whittaker index is 2.1 times higher,
the Morisita–Horn index is 7.5 times higher, and the Jaccard similarity index is 7.7 times
higher. Significant statistical differences (Mann–Whitney U–test) were detected for average
number of fungal species per plot (U = 0, Z = –3.36, p= 0.0008) between the greenhouses and
old-growth forest, as well as for Jaccard Index (U = 0, Z = –3.48,
p= 0.000
07). The Shannon
Index differed significantly between greenhouses and old-growth forests (
p< 0.01
), with
higher diversity in old-growth forests compared to the greenhouses.
J. Fungi 2023,9, 1116 16 of 23
Along the anthropogenic gradient from natural forests to the greenhouses, where the
proportion of non-native species of flora increased from 0% to 100%, and the average height
of stands decreased from 13.1 m to 2.6 m, the number of non-native fungal species grew
from 0 to 8, while the proportion of non-native fungal species increased from 0% to 20.5%,
the density of non-native species grew from 0 species to 13.1 species/ha.
A comparison of the lists of fungal species in the five studied plots indicates that
suburban and old-growth forests are clustered together, forest plantations in the city center
are combined with the Botanical Garden, and the mycobiota in greenhouses is significantly
different (Figure 5).
Figure 5.
Similarity of the species composition of aphyllophoroid macrofungi in 5 plots located along
anthropogenic gradient in Ekaterinburg city, suburban forests and old-growth forests.
In 2022, 5748 plant specimens (above 0.5 m) were counted in the EKB greenhouse
complex, of which 2167 were woody plants and 3581 were herbaceous. The oldest plants
are over 110 years old, but young individuals predominate—a third of the total numbers
of plants are aged from 0 to 20 years (Figure 6A). For all plants, as well as separately for
woody and herbaceous plants, a strong positive relationship was found between the age
and the number of fungal specimens developing on them (R
2
= 0.99). In all cases, the
relationship was S-shaped (Figure 6B): no fungi were found on young plants (0–20 years
old), and some fungi were found on 1–3% of plants aged 30–50 years only. Further, on
plants aged 60–90 years, the proportion of plants increased exponentially from 4–11% to
74–86%. As a result, the dependence reached a plateau—the fruitbodies of aphyllophoroid
fungi were collected on all plants (100%) over 100 years old.
J. Fungi 2023,9, 1116 17 of 23
J. Fungi 2023, 9, x FOR PEER REVIEW 17 of 23
Figure 6. Number of non-native plant specimens (woody, herbaceous and total) in the Ekaterinburg
city greenhouses divided into decade groups by decades from the youngest plants of 0–20 years old
to the oldest ones of 100–110 years old (A); and the percentage of plant specimens, on which aphyl-
lophoroid fungi fruitbodies were found (B).
4. Discussion
4.1. Aphyllophoroid Fungi in Greenhouses of the Three Boreal Cities
In the greenhouses of three boreal cities, on living and dead parts of non-native plants,
local species of fungi predominate, including 46 species, which is almost 80% of the total
list. They also predominate in terms of the number of samples (127) accounting for 90.7% of
the total number of samples. There are significantly fewer non-native species—12, the
findings of which represent single specimens.
More specimens of fungi have been collected on woody plants. In SPB, out of 17 sam-
ples, 16 were collected on woody plants, in MSC, 32 out of 38 were collected on woody
plants, and in EKB, 65 out of 85 were collected on woody plants. As the sampling effort in-
creases, the proportion of species on woody versus herbaceous plants decreases from 16.0
in SPB to 5.3 in MSC and to 3.4 in EKB.
It is worth noting that in the greenhouses of Central Europe (Austria, Germany, Po-
land, and Czech Republic), the number of aphyllophoroid fungi species varies from three
to eight species only [6,7,10,11,14,16–20,39]. A number of species identified in these studies
were also collected on non-native plants and in greenhouses of boreal Europe (Bjerkandera
adusta, Rigidoporus lineatus, and Trechispora farinacea), while others were absent (Asterostro-
ma cervicolor, Henningsomyces candidus, etc.). On building structures, boxes and tubs, species
were also collected indicated in greenhouses in Eastern Europe: Antrodia vaillantii, Coni-
ophora puteana, Hyphodontia quercina, Laetiporus sulphureus, Schizophyllum commune, but there
are also a number of species not collected by us, for example, Dentipellis fragilis (Pers.)
Donk, Hericium coralloides (Scop.) Pers., Postia floriformis (Quél.) Jülich, Steccherinum
ochraceum (Pers.) S.F. Gray. In Czech greenhouses, the number of fungal species depends
on the greenhouse area and the study duration. On the other hand, in the Austrian and
Polish greenhouses, it was found that the identified number of fungal species over a period
of 3–4 years no longer increased with repeated studies, which could indicate the identifica-
tion of a complete list [10,11,19].
In contrast to the Central European greenhouses, our study did not reveal any corre-
lation between the number of fungal species in greenhouses and their area, which was due
to the fact that the greenhouses in SPB and MSC had the largest areas. However, the
greatest number of species was collected in EKB, because purposeful studies were carried
out here for several years. In our opinion, for the same reason, there is no correlation be-
tween the average age of the greenhouses. We also found a strong positive relationship
Figure 6.
Number of non-native plant specimens (woody, herbaceous and total) in the Ekaterinburg
city greenhouses divided into decade groups by decades from the youngest plants of 0–20 years
old to the oldest ones of 100–110 years old (
A
); and the percentage of plant specimens, on which
aphyllophoroid fungi fruitbodies were found (B).
4. Discussion
4.1. Aphyllophoroid Fungi in Greenhouses of the Three Boreal Cities
In the greenhouses of three boreal cities, on living and dead parts of non-native plants,
local species of fungi predominate, including 46 species, which is almost 80% of the total
list. They also predominate in terms of the number of samples (127) accounting for 90.7%
of the total number of samples. There are significantly fewer non-native species—12, the
findings of which represent single specimens.
More specimens of fungi have been collected on woody plants. In SPB, out of 1
7 samp
les,
16 were collected on woody plants, in MSC, 32 out of 38 were collected on woody plants, and
in EKB, 65 out of 85 were collected on woody plants. As the sampling effort increases, the
proportion of species on woody versus herbaceous plants decreases from 16.0 in SPB to 5.3 in
MSC and to 3.4 in EKB.
It is worth noting that in the greenhouses of Central Europe (Austria, Germany,
Poland, and Czech Republic), the number of aphyllophoroid fungi species varies from
three to eight species only [
6
,
7
,
10
,
11
,
14
,
16
–
20
,
39
]. A number of species identified in these
studies were also collected on non-native plants and in greenhouses of boreal Europe
(Bjerkandera adusta,Rigidoporus lineatus, and Trechispora farinacea), while others were absent
(Asterostroma cervicolor,Henningsomyces candidus, etc.). On building structures, boxes and
tubs, species were also collected indicated in greenhouses in Eastern Europe: Antrodia
vaillantii,Coniophora puteana,Hyphodontia quercina,Laetiporus sulphureus,Schizophyllum
commune, but there are also a number of species not collected by us, for example, Dentipellis
fragilis (Pers.) Donk, Hericium coralloides (Scop.) Pers., Postia floriformis (Quél.) Jülich,
Steccherinum ochraceum (Pers.) S.F. Gray. In Czech greenhouses, the number of fungal
species depends on the greenhouse area and the study duration. On the other hand, in the
Austrian and Polish greenhouses, it was found that the identified number of fungal species
over a period of 3–4 years no longer increased with repeated studies, which could indicate
the identification of a complete list [10,11,19].
In contrast to the Central European greenhouses, our study did not reveal any correla-
tion between the number of fungal species in greenhouses and their area, which was due to
the fact that the greenhouses in SPB and MSC had the largest areas. However, the greatest
number of species was collected in EKB, because purposeful studies were carried out here
for several years. In our opinion, for the same reason, there is no correlation between the
average age of the greenhouses. We also found a strong positive relationship between
fungal species richness and plant age, where the strength increased when studying plants
aged 70 years and older (Figure 6).
J. Fungi 2023,9, 1116 18 of 23
Widespread local species of aphyllophoroid fungi predominate in greenhouses, but
Red Data Book and rare species do not develop. We did not find in the studied green-
houses any fungi included in the Federal Red Book of Russia [
48
] or in the Red Lists of
the three studied regions [
49
–
51
]. Only one protected species was collected in the EKB
greenhouses—the poroid Abortiporus biennis—which had an IUCN threat category (CR
B1; D1) in Sverdlovsk province [
31
]. This fungus grew in a woody tub at the base of the
Madagascar palm Dypsis lutescens (H. Wendl.) Beentje and J. Dransf.
Some non-native species of fungi in greenhouses are predominantly distributed in
the sub/tropical zones of the planet: Flaviporus brownii,Hymenochaete rheicolor,Lindtneria
trachyspora,Physalacria orientalis, and Rigidoporus lineatus; others are common in European
nemoral forests: Hypochniciellum ovoideum, and Thanatephorus terrigenus; but four are com-
mon in East Asia: Clavulina ornatipes,Clavulinopsis aurantiocinnabarina,Microporus xanthopus,
and Physalacria cryptomeriae. It is possible that this distribution of new species reflects the
centers, from which the plants were brought to the greenhouses. We know that in the early
1950s, some plant seedlings were brought to the Botanical Garden UrB RAS (EKB) along
with soil from the Russian Far East, which can explain the findings of East Asian fungal
species here. Each of the non-native species is collected in one city only. The proportion of
non-native species in greenhouses has a strongly positive relation to the number of fungal
species identified and selective effort. Undoubtedly, there are still few studied points to
prove this result, but for the future, it will be interesting to test on more material. Is this
dependence characteristic of aphyllophoroid fungi in the boreal zone only, or does it also
work in the nemoral zone?
In the three cities, only four species of pathogenic fungi were collected, which
was unexpected considering the old age of many plants as well as the studied group
of macrofungi—aphyllophoroids, which was the most common and numerous of phy-
topathogens in boreal forests [
8
,
27
]. It should be noted that pathogenic species of
agaricoid fungi are absent in the greenhouses of Central Europe [
7
,
10
,
11
,
19
]. This is due
to the regular fungicidal treatment of plants and the removal of diseased plants or dead
plant parts. All pathogens (including facultative) collected in greenhouses in Eastern
Europe (Athelia rolfsii,Helicobasidium purpureum,Thelephora palmata, and Th. terrestris)
do not cause dangerous diseases in adult plants, although some species from the genus
Thelephora can cause mass death of young seedlings of coniferous plants [
25
]. Three
of the