See discussions, stats, and author profiles for this publication at: https://www.researchgate.

net/publication/328098984

Importation of psychrotolerant fungi to Antarctica associated with wooden

cargo packaging

Article in Antarctic Science · October 2018

DOI: 10.1017/S0954102018000329

CITATIONS READS

10 214

4 authors:

Kevin A Hughes Marta Misiak

British Antarctic Survey Cardiff University
156 PUBLICATIONS 7,316 CITATIONS 3 PUBLICATIONS 31 CITATIONS

SEE PROFILE SEE PROFILE

Yogabaanu Ulaganathan Kevin K Newsham

Universiti Kebangsaan Malaysia British Antarctic Survey
2 PUBLICATIONS 41 CITATIONS 184 PUBLICATIONS 7,509 CITATIONS

SEE PROFILE SEE PROFILE

All content following this page was uploaded by Yogabaanu Ulaganathan on 02 May 2021.

The user has requested enhancement of the downloaded file.

 Antarctic Science 30(5), 298–305 (2018) © Antarctic Science Ltd 2018 doi:10.1017/S0954102018000329

Importation of psychrotolerant fungi to Antarctica associated with

wooden cargo packaging
KEVIN A. HUGHES1, MARTA MISIAK1, YOGABAANU ULAGANATHAN2 and KEVIN K. NEWSHAM1
1
British Antarctic Survey, NERC, High Cross, Madingley Road, Cambridge CB3 0ET, UK
2
National Antarctic Research Centre, University of Malaya, 50603 Lembah Pantai, Kuala Lumpur, Malaysia
kehu@bas.ac.uk

Abstract: The harsh climatic conditions and low levels of human activity in Antarctica, relative to other
regions, means few non-native species have established. However, the risk of introductions is becoming
greater as human activity increases. Non-native microorganisms can be imported to Antarctica in
association with fresh food, cargo and personal clothing, but the likelihood of their establishment is not
well understood. In January 2015, a wooden packing crate, heavily contaminated with fungi, was
imported by aircraft from Punta Arenas, Chile, to Rothera Research Station, Antarctica. Mucor
racemosus Bull. and two strains of Trichoderma viridescens (A.S. Horne & H.S. Will.) Jaklitsch &
Samuels were isolated from the wood. Measurements of hyphal extension rates indicated that all three
strains were psychrotolerant and capable of growth at 4°C, with M. racemosus growing at 0°C. The
imported fungi could grow at rates equivalent to, or faster than, species isolated from Antarctic soils,
suggesting that low temperature may not be a limiting factor for establishment. It is recommended that
wood heat-treatment standards, equivalent to those described in the International Standards for
Phytosanitary Measures No. 15, are employed by national operators importing cargo into Antarctica,
and that treated wood is adequately stored to prevent fungal contamination prior to transportation.
Received 22 February 2018, accepted 10 July 2018

Key words: Antarctic Treaty area, human impact, ISPM 15, microorganism, non-native, timber

Introduction probably transferred to the island on fresh vegetables,

Antarctica is geographically isolated and experiences
extreme climatic conditions compared to lower latitudes
(Laws 1984). In Antarctic terrestrial and freshwater
ecosystems, microorganisms (i.e. bacteria, algae, fungi
and viruses) may comprise the majority of the biomass
and biodiversity, and this may be the case particularly in
habitats subject to more climatically extreme conditions,
where higher organisms do not survive (Cowan & Ah
Tow 2004, Yergeau et al. 2007). Of the c. 1000 fungal
species reported from the Antarctic and sub-Antarctic
region, only 2–3% are considered to be endemic (Bridge &
Hughes 2010, https://legacy.bas.ac.uk/bas_research/data/
access/fungi/Speciespublic2.html accessed 26 December
2017), but research suggests endemism in microorganisms
may be greater and more widespread than thought
previously (Lawley et al. 2004, De Wever et al. 2009,
Vyverman et al. 2010). Antarctic communities may be at
risk if fungal strains are introduced from outside
Antarctica or moved between different Antarctic regions
(Cowan et al. 2011, Hughes et al. 2015a). Introduced
fungi may compete with native Antarctic species, alter
existing nutrient cycling or cause disease in native flora
and fauna (Bridge & Hughes 2010, Augustyniuk-Kram
2013). For example, on sub-Antarctic Marion Island, the
non-native Botrytis cinerea Pers. 1797, which was
now infects Kerguelen cabbage (Kloppers & Smith 1998).
Fungi and other microbes are likely to be transferred to
Antarctica by natural means, for example on wind
currents or on ocean debris and feathers of birds
(Marshall 1998, Hughes & Convey 2010, 2012), with
strains that are able to grow in the Antarctic natural
environment potentially becoming established.
Alternatively, fungi may be introduced to Antarctica by
humans (e.g. Wicklow 1968, Bridge & Hughes 2010,
Osyczka et al. 2012), with, as yet, little known
consequences for endemic microbial communities
(Vyverman et al. 2010).
The Antarctic continent has been subject to human
visitation for less than 200 years, but the extent of
national Antarctic operator and tourism industry activity
is increasing, resulting in greater risk of non-native species
introductions (Frenot et al. 2005, Tin et al. 2009, Hughes
et al. 2011a, 2015b). While research attention has focused
on the importation of plant propagules and invertebrates
(Whinam et al. 2005, Lee & Chown 2009, Hughes et al.
2010a, 2010b, Chown et al. 2012, Lityńska-Zając et al. 2012,
Tsujimoto & Imura 2012, Chwedorzewska et al. 2013,
Huiskes et al. 2014), less attention has been paid to the
threat of importing non-native microorganisms to
Antarctica (Broady & Smith 1994, Cowan et al. 2011,
Augustyniuk-Kram 2013) and in particular the risk

298

Downloaded from https://www.cambridge.org/core. NERC Library Service, on 08 Oct 2018 at 08:30:58, subject to the Cambridge Core terms of use, available at https://www.cambridge.org/core/terms.
https://doi.org/10.1017/S0954102018000329
 INTRODUCTION OF FUNGI TO ANTARCTICA IN WOOD
associated with contaminated timber packing materials
(Osyczka 2010, Osyczka et al. 2012).
This paper reports the unintentional importation of
fungi to Rothera Research Station, Antarctica, in
association with softwood cargo packaging. The hyphal
extension rates of the imported fungi at different
temperatures are examined, and the potential for their
establishment in Antarctica is discussed.
Methods
In January 2015, an item of cargo contained in a softwood
crate was flown from Punta Arenas, Chile, to the British
Antarctic Survey’s Rothera Research Station, Adelaide
Island, Antarctic Peninsula (Fig. 1). In general, the British
Antarctic Survey specifies the use of wood for packaging
that conforms to the International Standards on
Phytosanitary Measures 15 (ISPM 15), and this is
routinely used for cargo originating from the UK.
However, in this case the cargo was sourced and
packaged in Chile, and normal packing practices were
not observed. The bark-free wood that was used to
produce the bespoke crate was heavily contaminated with
fungal hyphae prior to construction of the crate, possibly
as a result of inappropriate storage practices. During the
six-hour flight it was observed that white and darkly
pigmented mycelia were present on the outer surfaces of
Fig. 1. Map of the Antarctic Peninsula and southern South
America, showing the transfer route of the contaminated
crate from Punta Arenas to Rothera Research Station.
Downloaded from https://www.cambridge.org/core. NERC Library Service, on 08 Oct 2018 at 08:30:58, subject to the Cambridge Core terms of use, available at https://www.cambridge.org/core/terms.
https://doi.org/10.1017/S0954102018000329
299
the packaging crate, except where it had been freshly
sawn during construction (Fig. 2). Darkly pigmented
fungal structures were also visible on the plank surfaces.
Upon arrival at Rothera Research Station, the station
management was informed of the importation of the
contaminated crate. The crate was off-loaded from
the aircraft, the catering equipment unpacked and the
packing crate quarantined. Following sampling, the
wooden packing crate was incinerated.
Fungal isolation
Wood fragments were sampled aseptically from below the
surface of the wooden crate and placed in sterile
polyethylene bags. In the laboratory, the wood
fragments were cut into small fragments (c. 1 × 2 mm)
aseptically and washed for 5 min in 10 changes of sterile
distilled water. Individual wood fragments were placed
onto potato dextrose agar medium (PDA; Oxoid;
Unipath Ltd, Basingstoke, UK) and incubated at 15°C.
Single fungal colonies were isolated after five days.
Fig. 2. Fungal-contaminated packing crate aboard the British
Antarctic Survey’s De Havilland DASH 7 en route to
Rothera Research Station, Antarctica, from Punta Arenas,
Chile. Areas of the wood colonized by dark pigmented
mycelia are visible on the plank surfaces (indicated by white
arrows).
 300
Following isolation, three strains were grown routinely
on PDA in 90 mm diameter Petri dishes.
Molecular identification
Fungal cultures were maintained on PDA medium at 4°C
for 10 weeks while in transit back to the UK from
Antarctica. Fresh cultures were prepared for DNA
extractions on half-strength PDA and incubated at 15°C.
A DNeasy Power Soil Kit (Qiagen, Manchester, UK) was
used to extract genomic DNA from the fungal cultures,
following the manufacturer’s protocol. DNA was extracted
from c. 0.3–0.5 g of fungal colony. The DNA extracts were
stored at -20°C prior to polymerase chain reaction (PCR)
amplification. Internal transcribed spacer (ITS) regions
of ribosomal RNA genes were amplified using ITS1F and
ITS4 primers (White et al. 1990, Gardes & Bruns 1993).
Polymerase chain reactions consisted of 4 µl of 5 × PCRBIO
Reaction Buffer (PCR Biosystems, London, UK), 0.2 µl of
each primer (20 pmol), 1 µl of template DNA and 0.5 U of
PCRBIO Taq DNA polymerase (PCR Biosystems, London,
UK), diluted with PCR grade water to a final volume of
20 µl. The PCR reactions were run in a PCRmax
thermocycler (AlphaCycler 4, PCRmax, Stone, UK) for 35
cycles, with each cycle consisting of 95°C for 30 s, 56°C for
30 s and 72°C for 30 s, with an initial denaturation step of 95°
C for 5 min and a final extension step at 72°C for 5 min. The
PCR products were separated by agarose gel electrophoresis
and visualized after staining with GelRed.
The PCR products were bidirectionally Sanger sequenced
(Source Bioscience, Cambridge, UK). Forward and reverse
chromatograms were edited and assembled, and primer
sequences trimmed using Geneious v. 6.1.6. The resulting
consensus ITS sequences were deposited in GenBank under
accession numbers MH279746 and MH279747. The
sequences were subjected to blastn searches against the
National Center for Biotechnology Information (NCBI)
nucleotide collection, with closest named matches being noted.
Growth rate analysis at different temperatures
Agar medium plugs taken from the colony margins of all
three fungi were inoculated onto PDA and incubated in
the dark at 0, 4, 15 and 25°C. Hyphal extension rate
analyses were carried out on three replicate Petri dishes of
each fungus per treatment, with a mean value per dish
being derived from three measurements made with sliding
calipers to an accuracy of 0.1 mm. Hyphal extension was
measured at regular intervals for up to 38 days and
extension rates calculated by linear regression.
Air temperature data
Mean monthly air temperature data were obtained for
Punta Arenas using World Meteorological Organization
Downloaded from https://www.cambridge.org/core. NERC Library Service, on 08 Oct 2018 at 08:30:58, subject to the Cambridge Core terms of use, available at https://www.cambridge.org/core/terms.
https://doi.org/10.1017/S0954102018000329
KEVIN A. HUGHES et al.
CLIMAT data (https://www.wmo.int/cpdb/chile), and
data for Rothera Research Station were obtained from
the SCAR READER database (https://www.scar.org/data-
products/ref-data-environmental-research/). Temperatures
in moss beds on Anchorage Island, Ryder Bay (67°36'18"S,
68°12'20"W), 5 km from Rothera Research Station, were
recorded with a Campbell CR10X Logger and IMKO
temperature probes.
Results
Strain identification
Three fungal strains were isolated from the wooden crate.
Molecular analyses showed that the ITS region sequences
of two isolates both matched at 100% identity and
100% coverage with that of Trichoderma viridescens
(A.S. Horne & H.S. Will.) Jaklitsch & Samuels reference
material (CBS 433.34), and the third matched at 99%
identity and 100% coverage with isolates of Mucor
racemosus Bull. (Table I).
Hyphal extension rate at different temperatures
Hyphal extension rate experiments showed differences in
the abilities of the three fungal isolates to grow at the
temperatures tested (Table II). All three strains grew at 4,
15 and 25°C. The two isolates of Trichoderma viridescens
displayed similar hyphal extension rates at 25°C and
15°C, but isolate A grew at over twice the rate of isolate B
at 4°C and was able to grow at a rate of 0.01 mm d-1 at
0°C, whereas isolate B did not grow at this temperature
(Table II). Mucor racemosus grew faster than either of
the T. viridescens isolates at all temperatures and was
able to grow at a rate of 1.25 mm d-1 at 0°C (Table II).
Air temperature data
Figure 3 shows the mean monthly air temperatures
recorded at Punta Arenas and Rothera Research Station
and the temperatures at 5 cm depth in moss on Anchorage
Island (5 km from Rothera Research Station). In general,
there was a difference in air temperature of c. 10°C
Table I. Identification of the three fungal strains isolated from the
imported softwood crate.
Coverage Identity GenBank
Isolate Species (%) (%) reference
A Trichoderma viridescens 100 100 NR_138429*
B Trichoderma viridescens 100 100 NR_138429*
C Mucor racemosus 100 99 KP411577
KP411551
KJ769665†
AJ878775†
*T. viridescens reference material (CBS 433.34); †f. sphaerosporus.
 INTRODUCTION OF FUNGI TO ANTARCTICA IN WOOD 301

Table II. Hyphal extension rates (mm d-1) at different temperatures of the three fungal strains imported in this study and fungi isolated from Antarctica in
other studies.
Temperature (°C)

Isolate Locations 25 17 15 4 0 -2 Reference

Trichoderma viridescens Crate transported to 12.84 − 9.29 8.64 0.01 − This study
(isolate A) Rothera Research (r2 = 0.997, − (r2 = 0.985, (r2 = 0.996, (r2 = 0.857,
Station n = 7) n = 7) n = 13) n = 12)
T. viridescens Crate transported to 12.56 − 9.28 3.26 0.00 − This study
(isolate B) Rothera Research (r2 = 0.987, − (r2 = 0.979, (r2 = 0.984, (r2 = NA,
Station n = 7) n = 7) n = 13) n = 12)
Mucor sp. Crate transported to 23.92 − 11.19 5.64 1.25 − This study
(isolate C) Rothera Research (r2 = 0.998, − (r2 = 0.988, (r2 = 0.984, (r2 = 0.924,
Station n = 7) n = 7) n = 9) n = 10)
Trichoderma sp. Rothera Research − 10.49 − 1.90 − − Hughes et al.
a
Station (2007)
Trichoderma koningii Rothera Research − 10.22 − 2.42 − − Hughes et al.
a
Station (2007)
Pythium sp. Rothera Point 5.69 − 5.20 0.65 − 0.06 Hughes et al.
b
(2003)
Phoma herbarum Rothera Point 1.26 − 1.36 0.62 − 0.05 Hughes et al.
b
(2003)
Mortierella parvispora Rothera Point 0.00 − 2.23 0.89 − 0.07 Hughes et al.
b
(2003)
Geomyces pannorum Rothera Point 0.02 − 0.52 0.20 − 0.05 Hughes et al.
b
(2003)
c
Geomyces pannorum Casey Station 0.84 − 0.98 0.63 − − Kerry (1990b)
(East Antarctica)
c
Mortierella sp. Casey Station 1.13 − 2.36 1.35 − − Kerry (1990b)
(East Antarctica)
c
Penicillium jensenii Casey Station 1.43 − 1.30 − − − Kerry (1990b)
(East Antarctica)
c
Phoma herbarum Casey Station 1.90 − 1.75 0.72 − − Kerry (1990b)
(East Antarctica)
c
Thelebolus microsporus Casey Station 0 − 1.44 1.00 − − Kerry (1990b)
(East Antarctica)
a
Fungi grown on minimal medium supplemented with glucose.
b
Fungi grown on potato dextrose agar.
c
Fungi grown on tomato agar and Lilly Barnett’s synthetic medium.

between Punta Arenas and Rothera Research Station
throughout the year, with winter mean monthly air
temperatures in Punta Arenas being roughly equivalent
to summer mean monthly air temperatures at Rothera
Research Station. However, mean monthly temperatures
of moss beds on Anchorage Island were up to 5°C higher
than mean air temperatures at nearby Rothera Research
Station.
Discussion
Three fungal stains were isolated from the cargo
packaging, although it is highly likely that other fungi
were also imported on the wood. Sequencing of ITS
regions of ribosomal RNA genes indicated that one of the
isolates was Mucor racemosus, the others being identified
as two strains of Trichoderma viridescens exhibiting
different hyphal extension rates at low (≤ 4°C)
temperatures. Trichoderma and Mucor are both
globally ubiquitous in soils and are thought to
contribute to the degradation of plant debris through
the production of extracellular enzymes but are
generally not able to degrade lignocelluloses (Eida
et al. 2011, Błaszczyk et al. 2016). Both genera have
been isolated previously from sub-Antarctic islands and
maritime and continental Antarctica (Corte & Daglio
1964, Sugiyama 1970, Hurst et al. 1983, Kerry 1984,
Fletcher et al. 1985, McRae & Seppelt 1999, Bradner
et al. 2000, Kochkina et al. 2012, Saili et al. 2014).
Trichoderma spp. were transported previously to
Antarctica in association with national operator
activities and detected in Antarctic soils subjected to
human impacts (Kerry 1990a, Hughes et al. 2007,
Osyczka et al. 2012), and, along with other fungi, were
also isolated from soils subjected to human activity near
Mawson, Davis and Rothera research stations, but were
not found locally in more pristine soils (Line 1988,
Hughes et al. 2007). Furthermore, both Trichoderma

Downloaded from https://www.cambridge.org/core. NERC Library Service, on 08 Oct 2018 at 08:30:58, subject to the Cambridge Core terms of use, available at https://www.cambridge.org/core/terms.
https://doi.org/10.1017/S0954102018000329
 302
Fig. 3. Mean monthly air temperature data for Punta Arenas,
Chile (1888–2017) (●) and Rothera Research Station,
Antarctica (1977–2017) (▼), and temperature recorded at
5 cm depth in moss at Anchorage Island, Ryder Bay, located
5 km from Rothera Research Station (□).
and Mucor spp. have been reported in studies
examining non-native fungi introduced to research
stations on clothing and equipment (Augustyniuk-
Kram 2013) and fresh foods (Hughes et al. 2011b),
with a Trichoderma strain also having been isolated
previously from wood packaging (Kerry 1990a).
Moreover, the high isolation frequency of Mucor spp.
and other fungal genera near Mawson Station (East
Antarctica) could have been due to introductions from
temperate regions in the time since the station was
established in 1954 (Kerry 1990a).
Previous studies have also suggested that fungi other
than Trichoderma and Mucor spp. may have been
introduced to the Antarctic. For example, introduced
wood decay fungi have been isolated from timber on
Deception Island, South Shetland Islands (Held &
Blanchette 2017), and Phialophora fastigiata Lagerb. &
Melin, potentially introduced on softwood packing
crates, has been found in contaminated soils around
Australian Antarctic research stations (Kerry 1990a,
Bölter et al. 2002). It has also been suggested that
Aspergillus fumigatus Fresen., 1863, which causes
aspergillosis in birds, was introduced to near an Adélie
penguin Pygoscelis adeliae (Hombron & Jacquinot)
colony at Cape Hallett in Victoria Land during the
period in which Hallett Station was present (Wicklow
1968, Bridge & Hughes 2010). Osyczka et al. (2012)
identified 13 ascomycetes and basidiomycetes imported to
Arctowski Station (King George Island, South Shetland
Islands) associated with timber, but suggested, without
confirmatory checks, that the probability of the
imported fungi becoming established and spreading
was rather negligible. However, research by Farrell et al.
(2011), who examined introduced and indigenous fungi
Downloaded from https://www.cambridge.org/core. NERC Library Service, on 08 Oct 2018 at 08:30:58, subject to the Cambridge Core terms of use, available at https://www.cambridge.org/core/terms.
https://doi.org/10.1017/S0954102018000329
KEVIN A. HUGHES et al.
of the Ross Island historic huts and pristine areas of
Antarctica, found that although non-Antarctic fungal
species were almost certainly introduced to the continent
in association with wood and other organic materials,
it was indigenous soil fungi that exploited these
substrates for nutrients. In contrast, Held & Blanchette
(2017) studied wood rot fungi in timber buildings on
Deception Island and suggested that brown and white
rot Basidiomycota were introduced with the wood and
building materials and were thriving under maritime
Antarctic conditions.
What is the likelihood of introduced fungi becoming
established in the Antarctic natural environment? In this
study of hyphal extension rates, all of the isolated fungi
exhibited psychrotolerance, with all three strains growing
at 4°C and Mucor racemosus showing relatively rapid
growth at 0°C. Mean summer soil temperature measured
at Anchorage Island (5 km from Rothera Point) was
reported as 2.8°C at 5 cm soil depth (Yergeau et al. 2012),
which is close to the January mean temperature reported
here of 4.3°C in moss at the same depth (Fig. 3).
Furthermore, it has been reported that coastal Antarctic
soils can attain summer temperatures of >15°C when
subjected to high levels of solar radiation (Smith 1988,
Davey et al. 1992). Therefore, soil temperature may not
be a limiting factor for the establishment of some of the
species imported to Rothera Research Station, although
other factors, such as salinity, water availability and
UV radiation, may have an influence (Kerry 1990b,
Arnold et al. 2003, Hughes et al. 2003, Ruisi et al. 2007).
The two Trichoderma viridescens strains isolated in this
study had faster hyphal extension rates at 4°C than two
Trichoderma strains isolated previously from
contaminated soils at Rothera Research Station
(Hughes et al. 2007), as well as other strains isolated
from pristine vegetated sites near Rothera Research
Station (Hughes et al. 2003) and Casey Station (Wilkes
Land, coastal East Antarctica; 64°17'S, 100o32'E) (Kerry
1990b) (Table II). Given that the three strains studied here
originated from Punta Arenas (53°10'S, 70°56-W), which
experiences mean winter (June–August) temperatures of
2.1°C (Fig. 3), it is unsurprising that the fungi showed
some pre-adaptation to growth at low temperatures. As
the physiologies of the three introduced fungal strains
studied here are apparently compatible with the Antarctic
summer climate, it is possible that they might have
become established in soil and other substrates close to
Rothera Research Station had steps not been taken to
destroy the packaging on which they were introduced
to the Antarctic. However, detecting the presence of
introduced microbes in the Antarctic natural
environment represents a considerable challenge. It is
possible that sequencing of ITS regions of precursor
ribosomal RNA (Rajala et al. 2011), a much shorter-lived
molecule than DNA that is predominantly present in
 INTRODUCTION OF FUNGI TO ANTARCTICA IN WOOD
metabolically active tissue, could be used to identify
whether introduced, non-endemic fungi are present in
Antarctic microbial communities.
Cargo packing material management
In 1991, the Antarctic Treaty Consultative Parties
(ATCPs) agreed Annex II to the Protocol on
Environmental Protection to the Antarctic Treaty
(entered into force in 1998), which stipulates that non-
native species are not to be intentionally introduced,
unless in accordance with a permit issued by an
appropriate national authority. More specifically, the
Protocol states that ‘each Party shall require that
precautions are taken to prevent the accidental
introduction of microorganisms (e.g. viruses, bacteria,
yeasts, fungi) not present naturally in the Antarctic
Treaty area’ and prohibits the deliberate introduction of
non-sterile soil.
All 29 Antarctic Treaty Consultative Parties
participate in the International Standards for
Phytosanitary Measures No. 15 (ISPM 15) developed by
the International Plant Protection Convention. The
Measure stipulates that wood materials of a thickness
greater than 6 mm, used to ship products between
countries, should be heat treated or fumigated to
prevent transfer of insects and disease-causing
microorganisms. In agreement with the 'Checklists for
supply chain managers of national Antarctic programmes
for the reduction in risk of transfer of non-native species'
(COMNAP & SCAR 2010), it is recommended that
equivalent wood heat-treatment standards are employed
by national operators importing cargo into Antarctica.
Furthermore, it is suggested that particular attention be
given to the adequate storage of wood to prevent fungal
contamination prior to transport into the Treaty area.
Antarctic microbial diversity may be impacted
irreversibly unless fuller consideration is given to the
routes and risks of microbial introductions and the
agreement of internationally applicable biosecurity
measures (Cowan et al. 2011, Convey et al. 2012,
Hughes et al. 2015a).
Acknowledgements
This paper is a contribution to the SCAR State of the
Antarctic Ecosystem (AntEco) research programme.
The authors are supported by NERC core funding to
the British Antarctic Survey’s Polar Science for Planet Earth
programmes 'Biodiversity, Evolution and Adaptation'
and 'Environment Office - Long Term Monitoring and
Survey' (EO-LTMS). The authors are grateful to Laura
Gerrish for map production, Alison Massey for technical
assistance, Will Goodall-Copestake for help with DNA
Downloaded from https://www.cambridge.org/core. NERC Library Service, on 08 Oct 2018 at 08:30:58, subject to the Cambridge Core terms of use, available at https://www.cambridge.org/core/terms.
https://doi.org/10.1017/S0954102018000329
303
sequence analyses, and to three anonymous reviewers,
who provided helpful comments on the manuscript.
Author contributions
KAH led the experimental work and the conceptual
development and writing of the paper. MM led the
molecular biological work. YU contributed to the
environmental sampling. KKN contributed expert
advice and helped with the drafting and revision of the
manuscript.
References
ARNOLD, R.J., CONVEY, P., HUGHES, K.A. & WYNN-WILLIAMS, D.D.
2003. Seasonal periodicity of physical factors, inorganic nutrients and
microalgae in Antarctic fellfields. Polar Biology, 26, 10.1007/s00300-
003-0503-2.
AUGUSTYNIUK-KRAM, A., CHWEDORZEWSKA, K.J., KORCZAK-ABSHIRE,
M., OLECH, M. & LITYŃSKA-ZAJĄC, M. 2013. An analysis of fungal
propagules transported to the Henryk Arctowski Station. Polish Polar
Research, 34, 10.2478/popore − 2013 − 0015.
BŁASZCZYK, L., STRAKOWSKA, J., CHEŁKOWSKI, J., GĄBKA-BUSZEK, A. &
KACZMAREK, J. 2016. Trichoderma species occurring on wood with
decay symptoms in mountain forests in Central Europe: genetic and
enzymatic characterization. Journal of Applied Genetics, 57, 10.1007/
s13353-015-0326-1.
BÖLTER, M., KANDELER, E., PIETR, S.J. & SEPPELT, R.D. 2002.
Heterotrophic microbes, microbial and enzymatic activity in
Antarctic soils. In BEYER, L. & BÖLTER, M., eds. Geoecology of
Antarctic ice-free coastal landscapes. Berlin: Springer, 189–214.
BRADNER, J.R., SIDHU, R.K., YEE, B., SKOTNICKI, M.L., SELKIRK, P.M.
& NEVALAINEN, K.M.H. 2000. A new microfungal isolate, Embellisia
sp., associated with the Antarctic moss Bryum argentum. Polar
Biology, 23, 10.1007/s003000000161.
BRIDGE, P.D. & HUGHES, K.A. 2010. Conservation issues for
Antarctic fungi. Mycologia Balcanica, 7, 11–14.
BROADY, P. & SMITH, R. 1994. A preliminary investigation of the
diversity, survivability and dispersal of algae introduced into
Antarctica by human activity. Proceedings of the NIPR Symposium
on Polar Biology, 7, 185–197.
CHOWN, S.L., HUISKES, A.H.L., GREMMEN, N.J.M. et al. 2012.
Continent-wide risk assessment for the establishment of
nonindigenous species in Antarctica. Proceeds of the National
Academy of Sciences USA, 109, 10.1073/pnas.1119787109.
CHWEDORZEWSKA, K.J., KORCZAK-ABSHIRE, M., OLECH, M., LITYŃSKA-
ZAJĄC, M. & AUGUSTYNIUK − KRAM, A. 2013. Alien invertebrates
transported accidentally to the Polish Antarctic Station in cargo and
on fresh foods. Polish Polar Research, 34, 55–66.
COMNAP & SCAR 2010. Checklists for supply chain managers of
national Antarctic programmes for the reduction in risk of transfer of
non-native species. Available at https://www.comnap.aq/SitePages/
checklists.aspx.
CONVEY, P., HUGHES, K.A. & TIN, T. 2012. Continental governance and
environmental management mechanisms under the Antarctic Treaty
System: sufficient for the biodiversity challenges of this century?
Biodiversity, 13, 10.1080/14888386.2012.703551.
CORTE, A. & DAGLIO, C.A.N. 1964. A mycological study of the
Antarctic air. In CARRICK, R., HOLDGATE, M.W. & PREVOST, J., eds.
Biologie Antarctique. Paris: Hermann, 115–120.
COWAN, D. A. & AH TOW, L. 2004. Endangered Antarctic environments.
Annual Reviews in Microbiology, 58, 10.1146/annurev.micro.
57.030502.090811.
 304 KEVIN A. HUGHES et al.
COWAN, D.A., CHOWN, S.L., CONVEY, P., TUFFIN, M., HUGHES, K.A.,
POINTINGS, S. & VINCENT, W.F. 2011. Non-indigenous microorganisms
in the Antarctic: assessing the risks. Trends in Microbiology, 19,
10.1016/j.tim.2011.07.008.
DAVEY, M.C., PICKUP, J. & BLOCK, W. 1992. Temperature variation and
its biological significance in fellfield habitats on a maritime
Antarctic island. Antarctic Science, 4, 10.1017/S0954102092000567.
DE WEVER, A., LELIAERT, F., VERLEYEN, E., VANORMELINGEN, P., VAN
DER GUCHT, K., HODGSON, D.A., SABBE, K. & VYVERMAN, W. 2009.
Hidden levels of phylodiversity in Antarctic green algae: further
evidence for the existence of glacial refugia. Proceedings of the Royal
Society B – Biological Sciences, 276, 10.1098/rspb.2009.0994.
EIDA, M.F., NAGAOKA, T., WASAKI, J. & KOUNO, K. 2011. Evaluation of
cellulolytic and hemicellulolytic abilities of fungi isolated from coffee
residue and sawdust composts. Microbes and Environments, 26,
10.1264/jsme2.ME10210.
FARRELL, R.L., ARENZ, B.E., DUNCAN, S.M., HELD, B.W., JURGENS, J.A.
& BLANCHETTE, R.A. 2011. Introduced and indigenous fungi of the
Ross Island historic huts and pristine areas of Antarctica. Polar
Biology, 34, 10.1007/s00300-011-1060-8.
FLETCHER, L.D., KERRY, E.J. & WESTE, G.M. 1985. Microfungi of Mac.
Robertson and Enderby Lands, Antarctica. Polar Biology, 4, 10.1007/
BF00442904.
FRENOT, Y., CHOWN, S.L., WHINAM, J., SELKIRK, P., CONVEY, P.,
SKOTNICKI, M. & BERGSTROM, D. 2005. Biological invasions in the
Antarctic: extent, impacts and implications. Biological Reviews, 80,
10.1017/S1464793104006542.
GARDES, M. & BRUNS, T.D. 1993. ITS primers with enhanced
specificity for basidiomycetes – application to the identification of
mycorrhizae and rusts. Molecular Ecology, 2, 10.1111/j.1365-
294X.1993.tb00005.x.
HELD, B.W. & BLANCHETTE, R.A. 2017. Deception Island, Antarctica,
harbors a diverse assemblage of wood decay fungi. Fungal Biology, 12,
10.1016/j.funbio.2016.11.009.
HUGHES, K.A. & CONVEY, P. 2010. The protection of Antarctic
terrestrial ecosystems from inter- and intra-continental transfer of
non-indigenous species by human activities: a review of current
systems and practices. Global Environmental Change, 20, 10.1016/j.
gloenvcha.2009.09.005.
HUGHES, K.A. & CONVEY, P. 2012. Determining the native/non-native
status of newly discovered terrestrial and freshwater species in
Antarctica – current knowledge, methodology and management
action. Journal of Environmental Management, 93, 10.1016/j.
jenvman.2011.08.017.
HUGHES, K.A., BRIDGE, P. & CLARK, M.S. 2007. Tolerance of Antarctic
soil fungi to hydrocarbons. Science of the Total Environment, 372,
10.1016/j.scitotenv.2006.09.016.
HUGHES, K.A., COWAN, D.A. & WILMOTTE, A. 2015a. Protection of
Antarctic microbial communities – ‘out of sight, out of mind’.
Frontiers in Microbiology, 10.3389/fmicb.2015.00151.
HUGHES, K.A., LAWLEY, B. & NEWSHAM, K.K. 2003. Solar UV-B
radiation inhibits the growth of Antarctic terrestrial fungi. Applied and
Environmental Microbiology, 69, 10.1128/AEM.69.3.1488-1491.200.
HUGHES, K.A., CONVEY, P., MASLEN, N.R. & SMITH, R.I.L. 2010a.
Accidental transfer of non-native soil organisms into Antarctica on
construction vehicles. Biological Invasions, 12, 10.1007/s10530-009-
9508-2.
HUGHES, K.A., PERTIERRA, L.R., MOLINA-MONTENEGRO, M. & CONVEY,
P. 2015b. Biological invasions in terrestrial Antarctica: what is the
current status and can we respond? Biodiversity and Conservation, 24,
10.1007/s10531-015-0896-6.
HUGHES, K.A., FRETWELL, P., RAE, J., HOLMES, K. & FLEMING, A.
2011a. Untouched Antarctica: mapping a finite and diminishing
environmental resource. Antarctic Science, 23, 10.1017/
S095410201100037X.
Downloaded from https://www.cambridge.org/core. NERC Library Service, on 08 Oct 2018 at 08:30:58, subject to the Cambridge Core terms of use, available at https://www.cambridge.org/core/terms.
https://doi.org/10.1017/S0954102018000329
HUGHES, K.A., LEE, J.E., WARE, C., KIEFER, K. & BERGSTROM, D.M.
2010b. Impact of anthropogenic transportation to Antarctica on alien
seed viability. Polar Biology, 33, 10.1007/s00300-010-0801-4.
HUGHES, K.A., LEE, J.E., TSUJIMOTO, M. et al. 2011b. Food for thought:
risks of non-native species transfer to the Antarctic region with fresh
produce. Biological Conservation, 144, 10.1016/j.biocon.2011.03.001.
HUISKES, A.H.L., GREMMEN, N.J.M., BERGSTROM, D.M. et al. 2014.
Aliens in Antarctica: assessing transfer of plant propagules by human
visitors to reduce invasion risk. Biological Conservation, 171, 10.1016/
j.biocon.2014.01.038.
HURST, J.L., PUGH, G.J.F. & WALTON, D.W.H. 1983. Fungal succession
and substrate utilisation on the leaves of three South Georgia
phanerograms. BAS Bulletin, No. 58. 89–100.
KERRY, E. 1984. The fungal flora of Macquarie Island. Tasmanian
Naturalist, 78, 16–21.
KERRY, E. 1990a. Microorganisms colonizing plants and soil subjected to
different degrees of human activity, including petroleum contamination,
in the Vestfold Hills and MacRobertson Land, Antarctica. Polar
Biology, 10, 10.1007/BF00233690.
KERRY, E. 1990b. Effects of temperature on growth rates of fungi from
subantarctic Macquarie Island and Casey, Antarctica. Polar Biology,
10, 10.1007/BF00238428.
KLOPPERS, F.J. & SMITH, V.R. 1998. First report of Botryotinia fuckeliana
on Kerguelen cabbage on the sub-Antarctic Marion Island. Plant
Disease, 82, 10.1094/PDIS.1998.82.6.710A.
KOCHKINA, G., IVANUSHKINA, N., OZERSKAYA, S., CHIGINEVA, N.,
VASILENKO, O., FIRSOV, S., SPIRINA, E. & GILICHINSKY, D. 2012.
Ancient fungi in Antarctic permafrost environments. FEMS
Microbiology Ecology, 82, 10.1111/j.1574-6941.2012.01442.x.
LAWLEY, B., RIPLEY, S., BRIDGE, P. & CONVEY, P. 2004. Molecular analysis
of geographic patterns of eukaryotic diversity in Antarctic soils.
Applied and Environmental Microbiology, 70, 10.1128/AEM.70.10.5963-
5972.2004.
LAWS, R.M. 1984. Antarctic ecology. London: Academic Press, 858 pp.
LEE, J.E. & CHOWN, S.L. 2009. Breaching the dispersal barrier to
invasion: quantification and management. Ecological Applications,
19, 10.1890/08-2157.1.
LINE, M.A. 1988. Microbial flora of some soils of Mawson Base and the
Vestfold Hills, Antarctica. Polar Biology, 8, 10.1007/BF00264718.
LITYNSKA-ZAJĄC, M., CHWEDORZEWSKA, K., OLECH, M., KORCZAK-
ABSHIRE, M. & AUGUSTYNIUK-KRAM, A. 2012. Diaspores and phyto-
remains accidentally transported to the Antarctic Station during three
expeditions. Biodiversity and Conservation, 21, 10.1007/s10531-012-
0371-6.
MARSHALL, W.A. 1998. Aerial transport of keratinaceous substrate and
distribution of the fungus Geomyces pannorum in Antarctic soils.
Microbial Ecology, 36, 10.1007/s002489900108.
MCRAE, C.F. & SEPPELT, R.D. 1999. Filamentous fungi of the
Windmill Islands, continental Antarctica: effect of water content
in moss turves on fungal diversity. Polar Biology, 22, 10.1007/
s003000050434.
OSYCZKA, P. 2010. Alien lichens unintentionally transported to the
Arctowski Station (South Shetlands, Antarctica). Polar Biology, 33,
10.1007/s00300-010-0786-z.
OSYCZKA, P., MLECZKO, P., KARASINSKI, D. & CHLEBICKI, A. 2012.
Timber transported to Antarctica: a potential and undesirable carrier
for alien fungi and insects. Biological Invasions, 14, 10.1007/s10530-
011-9991-0.
RAJALA, T., PELTONIEMI, M., HANTULA, J., MÄKIPÄÄ, R. & PENNANEN, T.
2011. RNA reveals a succession of active fungi during the decay of
Norway spruce logs. Fungal Ecology, 4, 10.1016/j.funeco.2011.05.
005.
RUISI, S., BARRECA, D., SELBMANN, L., ZUCCONI, L. & ONOFRI, S. 2007. Fungi
in Antarctica. Reviews in Environmental Science and Biotechnology,
6, 10.1007/s11157-006-9107-y.
 INTRODUCTION OF FUNGI TO ANTARCTICA IN WOOD
SAILI, N.S., SIDDIQUEE, S., VUI LING, C.M.W., GONZÁLEZ, M. & VIJAY
KUMAR, S. 2014. Lignocellulolytic activities among Trichoderma isolates
from Lahad Datu, Sabah and Deception Island, Antarctic. Journal of
Microbial Biochemistry and Technology, 6, 10.4172/1948-5948.1000159.
SMITH, R.I.L. 1988. Recording bryophyte microclimate in remote and
severe environments. In GLIME, J.M., ed. Methods in bryology.
Proceedings of the bryology methods workshop, Mainz. Nichinan,
Japan: Hattori Botanical Laboratory, 275–284.
SUGIYAMA, J. 1970. World’s last frontier III: polar mycology in
Antarctica. Polar News, 6, 17–24.
TIN, T., FLEMING, Z.L., HUGHES, K.A., AINLEY, D.G., CONVEY, P.,
MORENO, C.A., PFEIFFER, S., SCOTT, J. & SNAPE, I. 2009. Impacts of
local human activities on the Antarctic environment. Antarctic
Science, 21, 10.1017/S0954102009001722.
TSUJIMOTO, M. & IMURA, S. 2012. Does a new transportation system
increase the risk of importing non-native species to Antarctica?
Antarctic Science, 24, 10.1017/S0954102012000272.
VYVERMAN, W., VERLEYEN, E., WILMOTTE, A. et al. 2010. Evidence
for widespread endemism among Antarctic micro-organisms. Polar
Science, 4, 10.1016/j.polar.2010.03.006.
Downloaded from https://www.cambridge.org/core. NERC Library Service, on 08 Oct 2018 at 08:30:58, subject to the Cambridge Core terms of use, available at https://www.cambridge.org/core/terms.
https://doi.org/10.1017/S0954102018000329
View publication stats
305
WHINAM, J., CHILCOTT, N. & BERGSTROM, D.M. 2005. Subantarctic
hitchhikers: expeditioners as vectors for the introduction of alien
organisms. Biological Conservation, 121, 10.1016/j.biocon.2004.
04.020.
WHITE, T.J., BRUNS, T., LEE, S. & TAYLOR, J.W. 1990. Amplification and direct
sequencing of fungal ribosomal RNA genes for phylogenetics. In INNIS,
M.A., GELFAND, D.H., SNINSKY, J.J. & WHITE, T.J., eds. PCR protocols: a
guide to methods and applications. New York: Academic Press, 315–322.
WICKLOW, D.T. 1968. Aspergillus fumigatus Fresenius isolated from
ornithogenic soil collected at Hallett Station, Antarctica. Canadian
Journal of Microbiology, 14, 10.1139/m68-119.
YERGEAU, E., BOKHORST, S., HUISKES, A.H.L., BOSCHKER, H.T.S.,
AERTS, R. & KOWALCHUK, G.A. 2007. Size and structure of
bacterial, fungal and nematode communities along an Antarctic
environmental gradient. FEMS Microbial Ecology, 59, 10.1111/
j.1574-6941.2006.00200.x.
YERGEAU, E., BOKHORST, S., KANG, S., ZHOU, J.Z., GREER, C.W., AERTS,
R. & KOWALCHUK, G.A. 2012. Shifts in soil microorganisms in
response to warming are consistent across a range of Antarctic
environments. ISME Journal, 6, 10.1038/ismej.2011.124.